Rice blast is one of the important diseases in major rice producing areas of China. The main blast resistance genes Pi-ta and Pi-b showed broad-spectrum and durable resistance to rice blast in many rice growing areas ...Rice blast is one of the important diseases in major rice producing areas of China. The main blast resistance genes Pi-ta and Pi-b showed broad-spectrum and durable resistance to rice blast in many rice growing areas of China, which have been widely utilized in rice breeding and commercial production. In this study, on the basis of detection and verification of the genotypes of 22 rice varieties har- boring known blast resistance genes （Pi-ta and Pi-b） and blast susceptibility genes （pi-ta and pi-b）, two multiple PCR systems for these genes were established by us- ing the functional markers of blast resistance genes Pi-ta and Pi-b as well as blast susceptibility genes pi-ta and pi-b, respectively. Specifically, multiple PCR system I could simultaneously detect blast resistance genes Pi-ta and Pi-b, while system II could detect simultaneously blast susceptibility genes pi-ta and pi-b. In addition, the genotypes of 336 high generation breeding materials were detected with these two multiple PCR systems. The results were highly consistent with those of conventional single mark detection, indicating that these two multiplex PCR systems were stable, reliable and time-saving. The established multiplex PCR systems may serve as a rapid and efficient method to identify and screen rice germplasm resources and can be applied in marker-assisted selection to polymerize multiple genes for blast resis- tance in rice breeding.展开更多
AIM To observe the drug sensitizing effect andrelated mechanisms of fas gene transduction onhuman drug-resistant gastric cancer cellSGC7901/VCR(resistant to Vincristine).METHODS The cell cycle alteration wasobserved b...AIM To observe the drug sensitizing effect andrelated mechanisms of fas gene transduction onhuman drug-resistant gastric cancer cellSGC7901/VCR(resistant to Vincristine).METHODS The cell cycle alteration wasobserved by FACS.The sensitivity of gastriccancer cells to apoptosis was determined by invitro apoptosis assay.The drug sensitization ofcells to several anti-tumor drugs was observedby MTT assay.Immunochemical method wasused to show expression of P-gp and Topo Ⅱ ingastric cancer cells.RESULTS Comparing to SGC7901 and pBK-SGC7901/VCR,fas-SGC7901/VCR showeddecreasing G2 cells and increasing S cells,theG2 phase fraction of pBK-SGC7901/VCR wasabout 3.0 times that of fas-SGC7901/VCR,but Sphase fraction of fas-SGC7901/VCR was about1.9 times that of pBK-SGC7901/VCR,indicatingS phase arrest of fas-SGC7901/VCR.FACS alsosuggested apoptosis of fas-SGC7901/VCR,fas-SGC7901/VCR was more sensitive to apoptosisinducing agent VM-26 than pBK-SGC7901/VCR.MTT assay showed increased sensitization offas-SGC7901/VCR to DDP,MMC and 5-FU,butsame sensitization to VCR according to pBK-SGC7901/VCR.SGC7901,pBK-SGC7901/ VCRand fas-SGC7901/VCR had positively stainedTopo Ⅱ equally.P-gp staining in pBK- SGC7901/VCR was stronger than in SG07901,but there was little staining of P-gp in fas.SGC7901/VCR.CONCLUSION fas gene transduction couldreverse the MDR of human drug-resistant gastriccancer cell SGC7901/VCR to a degree,possiblybecause of higher sensitization to apoptosis anddecreased expression of P-gp.展开更多
Multiple myeloma(MM) is a cancer caused by uncontrolled proliferation of antibody-secreting plasma cells in bone marrow, which represents the second most common hematological malignancy. MM is a highly heterogeneous d...Multiple myeloma(MM) is a cancer caused by uncontrolled proliferation of antibody-secreting plasma cells in bone marrow, which represents the second most common hematological malignancy. MM is a highly heterogeneous disease and can be classified into a spectrum of subgroups based on their molecular and cytogenetic abnormalities. In the past decade, novel therapies, especially, the first-in-class proteasome inhibitor bortezomib, have been revolutionary for the treatment of MM patients. Despite these remarkable achievements, myeloma remains incurable with a high frequency of patients suffering from a relapse, due to drug resistance. Mutation in the proteasome β5-subunit(PSMB5) was found in a bortezomib-resistant cell line generated via long-term coculture with increasing concentrations of bortezomib in 2008, but their actual implication in drug resistance in the clinic has not been reported until recently. A recent study discovered four resistance-inducing PSMB5 mutations from a relapsed MM patient receiving prolonged bortezomib treatment. Analysis of the dynamic clonal evolution revealed that two subclones existed at the onset of disease, while the other two subclones were induced. Protein structural modeling and functional assays demonstrated that all four mutations impaired the binding of bortezomib to the 20 S proteasome, conferring different degrees of resistance. The authors further demonstrated two potential approaches to overcome drug resistance by using combination therapy for targeting proteolysis machinery independent of the 20 S proteasome.展开更多
Objective Antimicrobial resistance(AMR)has become a global concern and is especially severe in China.To effectively and reliably provide AMR data,we developed a new high-throughput real-time PCR assay based on microfl...Objective Antimicrobial resistance(AMR)has become a global concern and is especially severe in China.To effectively and reliably provide AMR data,we developed a new high-throughput real-time PCR assay based on microfluidic dynamic technology,and screened multiple AMR genes in broiler fecal samples.Methods A high-throughput real-time PCR system with an new designed integrated fluidic circuit assay were performed AMR gene detection.A total of 273 broiler fecal samples collected from two geographically separated farms were screened AMR genes.Results The new assay with limits of detection ranging from 40.9 to 8,000 copies/reaction.The sensitivity rate,specificity rate,positive predictive value,negative predictive value and correct indices were 99.30%,98.08%,95.31%,99.79%,and 0.9755,respectively.Utilizing this assay,we demonstrate that AMR genes are widely spread,with positive detection rates ranging from 0 to 97.07%in 273 broiler fecal samples.bla CTX-M,bla TEM,mcr-1,fex A,cfr,optr A,and int I1 showed over 80%prevalence.The dissemination of AMR genes was distinct between the two farms.Conclusions We successfully established a new high-throughput real-time PCR assay applicable to AMR gene surveillance from fecal samples.The widespread existence of AMR genes detected in broiler farms highlights the current and severe problem of AMR.展开更多
Clubroot and herbicide resistance,high oleic acid(OA)content,and early maturity are targets of rapeseed(Brassica napus L.)breeding.The objective of this study was to develop new male-fertility restorer lines by pyrami...Clubroot and herbicide resistance,high oleic acid(OA)content,and early maturity are targets of rapeseed(Brassica napus L.)breeding.The objective of this study was to develop new male-fertility restorer lines by pyramiding favorable genes to improve these traits simultaneously.Seven elite alleles for the four traits were introduced into the restorer line 621R by speed breeding with marker-assisted and phenotypic selection.Six introgression lines(ILs)were developed with four-to seven-gene combinations and crossed with two elite parents to develop hybrids.All ILs and their corresponding hybrids displayed high resistance to both clubroot pathotype 4 and sulfonylurea herbicides.Three ILs and their hybrids showed large increases in OA contents and four showed earlier maturity.These new ILs may be useful in rapeseed hybrid breeding for the target traits.展开更多
The introduction of Bacillus thuringiensis(Bt)cotton has reduced the burden of pests without harming the environment and human health.However,the efficacy of Bt cotton has decreased due to field-evolved resistance in ...The introduction of Bacillus thuringiensis(Bt)cotton has reduced the burden of pests without harming the environment and human health.However,the efficacy of Bt cotton has decreased due to field-evolved resistance in insect pests over time.In this review,we have discussed various factors that facilitate the evolution of resistance in cotton pests.Currently,different strategies like pyramided cotton expressing two or more distinct Bt toxin genes,refuge strategy,releasing of sterile insects,and gene silencing by RNAi are being used to control insect pests.Pyramided cotton has shown resistance against different cotton pests.The multiple genes pyramiding and silencing(MGPS)approach has been proposed for the management of cotton pests.The genome information of cotton pests is necessary for the development of MGPS-based cotton.The expression cassettes against various essential genes involved in defense,detoxification,digestion,and development of cotton pests will successfully obtain favorable agronomic characters for crop protection and production.The MGPS involves the construction of transformable artificial chromosomes,that can express multiple distinct Bt toxins and RNAi to knockdown various essential target genes to control pests.The evolution of resistance in cotton pests will be delayed or blocked by the synergistic action of high dose of Bt toxins and RNAi as well as compliance of refuge requirement.展开更多
In marker-assisted breeding for bacterial blight(BB) resistance in rice, three major resistance genes, viz., Xa21, xa13 and xa5, are routinely deployed either singly or in combinations. As efficient and functional mar...In marker-assisted breeding for bacterial blight(BB) resistance in rice, three major resistance genes, viz., Xa21, xa13 and xa5, are routinely deployed either singly or in combinations. As efficient and functional markers are yet to be developed for xa13 and xa5, we have developed simple PCR-based functional markers for both the genes. For xa13, we designed a functional PCR-based marker, xa13-prom targeting the In Del polymorphism in the promoter of candidate gene Os8N3 located on chromosome 8 of rice. With respect to xa5, a multiplex-PCR based functional marker system, named xa5 FM, consisting of two sets of primer pairs targeting the 2-bp functional nucleotide polymorphism in the exon II of the gene TFIIA5(candidate for xa5), has been developed. Both xa13-prom and xa5 FM can differentiate the resistant and susceptible alleles for xa13 and xa5, respectively, in a co-dominant fashion. Using these two functional markers along with the already reported functional PCR-based marker for Xa21(p TA248), we designed a single-tube multiplex PCR based assay for simultaneous detection of all the three major resistance genes and demonstrated the utility of the multiplex marker system in a segregating population.展开更多
Summary: The expression and functional activity of multiple drug resistance (MDR1) gene in human normal bone marrow CD34+ cells was observed. Human normal bone marrow CD34+ cells were enriched with magnetic cell sorti...Summary: The expression and functional activity of multiple drug resistance (MDR1) gene in human normal bone marrow CD34+ cells was observed. Human normal bone marrow CD34+ cells were enriched with magnetic cell sorting (MACS) system, and then liposome-mediated MDR1 gene was transferred into bone marrow CD34+ cells. Fluorescence-activated cell sorter was used to evaluate the expression and functional activity of P-glycoprotein (P-gp) encoded by MDR1 gene. It was found that the purity of bone marrow CD34+ cells was approximately (91±4.56) % and recovery rate was (72.3±2.36) % by MACS. The expression of P-gp in the transfected CD34+cells was obviously higher than that in non-transfected CD34+ cells. The amount of P-gp in non-transfected CD34+ cells was (11.2±2.2) %, but increased to (23.6±2.34) % 48 h after gene transfection (P<0.0l). The amount of P-gp was gradually decreased to the basic level one week later. The accumulation and extrusion assays showed that the overexpression of P-gp could efflux Rh-123 out of cells and there was low fluorescence within the transfected cells. The functional activity of P-gp could be inhibited by 10 μg/ml verapamil. It was suggested that the transient and highly effective expression and functional activity of P-gp could be obtained by liposome-mediated MRD1 transferring into human normal bone marrow CD34+ cells.展开更多
Multiple myeloma(MM)is a hematological tumor with high mortality and recurrence rate.Carfilzomib is a new-generation proteasome inhibitor that is used as the first-line therapy for MM.However,the development of drug r...Multiple myeloma(MM)is a hematological tumor with high mortality and recurrence rate.Carfilzomib is a new-generation proteasome inhibitor that is used as the first-line therapy for MM.However,the development of drug resistance is a pervasive obstacle to treating MM.Therefore,elucidating the drug resistance mechanisms is conducive to the formulation of novel therapeutic therapies.To elucidate the mechanisms of carfilzomib resistance,we retrieved the GSE78069 microarray dataset containing carfilzomib-resistant LP-1 MM cells and parental MM cells.Differential gene expression analyses revealed major alterations in the major histocompatibility complex(MHC)and cell adhesion molecules.The upregulation of the tumor necrosis factor(TNF)receptor superfamily member 1A(TNFRSF1A)gene was accompanied by the downregulation of MHC genes and cell adhesion molecules.Furthermore,to investigate the roles of these genes,we established a carfilzomib-resistant cell model and observed that carfilzomib resistance induced TNFRSF1A overexpression and TNFRSF1A silencing reversed carfilzomib resistance and reactivated the expression of cell adhesion molecules.Furthermore,TNFRSF1A silencing suppressed the tumorigenesis of MM cells in immunocompetent mice,indicating that TNFRSF1A may lead to carfilzomib resistance by dampening antitumor immunity.Furthermore,our results indicated that TNFRSF1A overexpression conferred carfilzomib resistance in MM cells and suppressed the expression of MHC genes and cell adhesion molecules.The suppression of MHC genes and cell adhesion molecules may impair the interaction between immune cells and cancer cells to impair antitumor immunity.Future studies are warranted to further investigate the signaling pathway underlying the regulatory role of TNFRSF1A in MM cells.展开更多
Rice yellow mottle virus (RYMV) is a major biotic constraint for rice production in Africa. The resistance-breaking ability of Tanzanian RYMV strains and phylotypes (S4lm (Tz526), S4lv (Tz516), S4ug (Tz508), S5 (Tz429...Rice yellow mottle virus (RYMV) is a major biotic constraint for rice production in Africa. The resistance-breaking ability of Tanzanian RYMV strains and phylotypes (S4lm (Tz526), S4lv (Tz516), S4ug (Tz508), S5 (Tz429, Tz445), S6c (Tz486) and S6w (Tz539)) were tested by inoculating rice cultivars with RYMV1 resistant alleles (Gigante (rymv1-2), Tog12387 (rymv1-3), Tog5681 (rymv1-3), Tog5438 (rymv1-4), Tog5672 (rymv1-4+rymv2) and Tog5674 (rymv 1-5)) in a screen house. The results revealed multiple resistance-breaking strains and phylotypes on resistant cultivars Gigante, Tog12387, Tog5438 and Tog5681. However, the resistance breakdown was highly variable depending on the strain used, and disease severity ranged from 11% - 75.3%. The virulence potential of RYMV phylotype S4lm (Tz526) was similar to phylotype S6w (Tz539). The impact of strains and phylotypes on yield and its components in rice cultivars revealed highly significant differences (P ≤ 0.001). The lowest percent plant height reduction (2.8%), number of tillers per plant (2.5%), 1000 grain weight (2.7%), spikelet sterility (3.5%) and yield (5%) was recorded in rice cultivar Gigante inoculated with RYMV phylotype S6c (Tz486). Phylotype S6c (Tz486) despite being less virulent compared to other strains, its virus titer in rice cultivar Gigante (1.833) was higher than S5 (Tz429, Tz445) inoculated on Tog5674 (0.171, 0.207) and S6w (Tz539) inoculated on Tog5681 (0.283). The resistant-breaking strain S5 (Tz445) multiplied in resistant rice cultivar Tog5674 without inducing visible symptoms but showed positive reaction to ELISA with low virus titer. The strain S5 overcame wide range of resistant alleles including rymv1-2, rymv1-3, rymv1-4 and rymv1-5 resistance, with exception of rymv1-4 + rymv2. The current results gave a new perspective for future identification of resistance-breaking mutations through sequencing of the RYMV genome in infected rice cultivars and mutagenesis of an infectious viral clone useful for future RYMV resistant breeding programs.展开更多
A multiplex PCR was developed to detect benzimidazole-resistance (BZ-R) or -susceptibility (BZ-S) in Haemonchus contortus by amplification with 4 primers of a sequence of the GRU-1 gene of β-tubulin of H. contortus m...A multiplex PCR was developed to detect benzimidazole-resistance (BZ-R) or -susceptibility (BZ-S) in Haemonchus contortus by amplification with 4 primers of a sequence of the GRU-1 gene of β-tubulin of H. contortus making use of sequence information available in Genbank. The method was based on two allele-non-specific primers and two allele- specific primers. F1 (264 bp) and F3 (799 bp) should be produced in BZ-R, F2 (585 bp) and F3 in BZ-S. With this method, we demonstrated that H. contortus BZ-R strain from Australia showed F1 and F3, and the worm BZ-S strain from Shanghai did F2 and F3. Sequence analysis of the isotype 1 gene of β-tubulin of BZ-R from Australia and BZ-S from Shanghai showed the code in residue 200 of the gene was respectively TAC and TTC. The LD50 of albendazole of the Australian BZ- R strain was 0.54 μg mL-1, the Shanghai BZ-S strain was only 0.0023 μg mL-1 by EHA (egg hatch assay). The multiplex PCR could determinate the genotype of single adult worm or several third stage larvae and was performed on at least 50 ng of genomic DNA. BZ-R H. contortus were not detected in Shihezi and Yining of the Xinjiang, Wuhe of the Anhui Province, Nanjing and Xuzhou of the Jiangsu Province. The LD50 of the H. contortus from these locations to albendazole as determined by EHA varied between 0.0023-0.0032 μg mL-1. The result indicated that the multiplex PCR could be used to differentiate BZ-R and BZ-S of H. contortus and that the BZ-R situation of H. contortus was not serious in China.展开更多
CRISPR homing gene drives have considerable potential for managing populations of medically and agriculturally significant insects.They operate by Cas9 cleavage followed by homology-directed repair,copying the drive a...CRISPR homing gene drives have considerable potential for managing populations of medically and agriculturally significant insects.They operate by Cas9 cleavage followed by homology-directed repair,copying the drive allele to the wild-type chromosome and thus increasing in frequency and spreading throughout a population.However,resistance alleles formed by end-joining repair pose a significant obstacle.To address this,we create a homing drive targeting the essential hairy gene in Drosophila melanogaster.Nonfunctional resistance alleles are recessive lethal,while drive carriers have a recoded“rescue”version of hairy.The drive inheritance rate is moderate,and multigenerational cage studies show drive spread to 96%–97%of the population.However,the drive does not reach 100%due to the formation of functional resistance alleles despite using four gRNAs.These alleles have a large deletion but likely utilize an alternate start codon.Thus,revised designs targeting more essential regions of a gene may be necessary to avoid such functional resistance.Replacement of the rescue element’s native 3'UTR with a homolog from another species increases drive inheritance by 13%–24%.This was possibly because of reduced homology between the rescue element and surrounding genomic DNA,which could also be an important design consideration for rescue gene drives.展开更多
In the present research,we proposed a scheme to address the issues of severe heat damage,high energy consumption,low cooling system efficiency,and wastage of cold capacity in mines.To elucidate the seasonal variations...In the present research,we proposed a scheme to address the issues of severe heat damage,high energy consumption,low cooling system efficiency,and wastage of cold capacity in mines.To elucidate the seasonal variations of environmental temperature through field measurements,we selected a high-temperature working face in a deep mine as our engineering background.To enhance the heat damage control cability of the working face and minimize unnecessary cooling capac-ity loss,we introduced the multi-dimensional heat hazard prevention and control method called"Heat source barrier and cooling equipment".First,we utilize shotcrete and liquid nitrogen injection to eliminate the heat source and implemented pressure equalization ventilation to disrupt the heat transfer path,thereby creating a heat barrier.Second,we establish divi-sional prediction models for airflow temperature based on the variation patterns obtained through numerical simulation.Third,we devise the location and dynamic control strategy for the cooling equipment based on the prediction models.The results of field application show that the heat resistance and cooling linkage method comply with the safety requirement throughout the entire mining cycle while effectively reducing energy consumption.The ambient temperature is maintained below 30℃,resulting in the energy saving of 10%during the high-temperature period and over 50%during the low-temperature period.These findings serve as a valuable reference for managing heat damage in high-temperature working faces.展开更多
Coastal lagoons provide many important services to human society,but their year-round use for aquaculture introduces large amounts of sewage.The contamination of antibiotic resistance genes(ARGs)is therefore of great ...Coastal lagoons provide many important services to human society,but their year-round use for aquaculture introduces large amounts of sewage.The contamination of antibiotic resistance genes(ARGs)is therefore of great concern.In this study,50 ARGs subtypes,two integrase genes(intl1,intl2),and 16S rRNA genes were detected by high-throughput quantitative PCR,and standard curves of all target genes were prepared for quantification.The occurrence and distribution of ARGs in a typical coastal lagoon(XinCun lagoon,China)were comprehensively explored.We detected 44 and 38 subtypes of ARGs in the water and sediment,respectively,and discuss the various factors influencing the fate of ARGs in the coastal lagoon.Macrolides-lincosamides-streptogramins B was the primary ARG type,and macB was the predominant subtype.Antibiotic efflux and antibiotic inactivation were the main ARG resistance mechanisms.The XinCun lagoon was divided into eight functional zones.The ARGs showed a distinct spatial distribution owing to the influence of microbial biomass and anthropogenic activity in different functional zones.Fishing rafts,abandoned fish ponds,the town sewage zone,and mangrove wetlands provided a large quantity of ARGs to the XinCun lagoon.Nutrients and heavy metals also significantly correlated with the fate of the ARGs,especially NO_(2)^(−)-N and Cu,which cannot be ignored.It is noteworthy that lagoon-barrier systems coupled with persistent pollutant inputs result in coastal lagoons acting as a“buffer pool”for ARGs,which can then accumulate and threaten the offshore environment.展开更多
Drug resistance is becoming a great problem in developing countries due to excessive use and misuse of antibiotics. The emergence of new pathogenic strains with resistance developed against most of the antibiotics whi...Drug resistance is becoming a great problem in developing countries due to excessive use and misuse of antibiotics. The emergence of new pathogenic strains with resistance developed against most of the antibiotics which may cause,difficult to treat infection.To understand the current scenario in different mode of infection is most important for the clinicians and medical practitioners.This article summarized some common infections and antibiotic resistance pattern found among these pathogens.展开更多
This study was to investigate the current trends of multiple drug resistance in bacteria against antibiotics for the proper empirical treatmen.Clinical isolates were collected from community-acquired infection of urin...This study was to investigate the current trends of multiple drug resistance in bacteria against antibiotics for the proper empirical treatmen.Clinical isolates were collected from community-acquired infection of urinary tract patients in Aligarh India from March 1999 to August 1999.Antibiotic susceptibility test was performed,using the disc diffusion method followed by plasmid isolation by the method of Kado and Liu.Transfer experiments were performed by the method of Lederberg and Cohen.Clinical study revealed that this infection was more common in young women.Various strains of E.coli isolated during the course of study were found to show multiple antibiotic resistance which was further characterized as plasmid-borne drug resistance.This study shows that E.coli may be one of the important causative agents of urinary tract infection(UTI) in young women.展开更多
Objective:To understand the insecticide resistance status of Culex quinquefasciatus Say（Diptera: Culicidae）（Cx.Quinquefasciatus） to deltamethrin,cyfluthrin,permethrin,lambdacyhalothrin, DDT and malathion in filari...Objective:To understand the insecticide resistance status of Culex quinquefasciatus Say（Diptera: Culicidae）（Cx.Quinquefasciatus） to deltamethrin,cyfluthrin,permethrin,lambdacyhalothrin, DDT and malathion in filarial endemic areas of Uttar Pradesh,India.Methods:Insecticide susceptibility assays were performed on wild-caught adult female Cx.quinquefasciatus mosquitoes to deltamethrin（0.05%）,cyfluthrin（0.15%）,permethrin（0.75%）,lambdacyhalothrin （0.05%）,malathion（5.0%） and DDT（4.0%）,the discriminating doses recommended by the World Health Organisation（WHO）.Results:The data showed that Cx.quinquefasciatus is highly resistant to DDT and malathion;the mortality was 28.33%and 27.5%,respectively and incipient resistance to synthetic pyrethroids（deltamethrin,cyfluthrin,permethrin,and lambdacyhalothrin）, where mortality ranged from 95.83%in permethrin to 98.33%in cyfluthrin and lambdacyhalothrin. Knockdown times(KDT<sub>50</sub>) in response to synthetic pyrethroids varied significantly between different insecticides（P【0.01） from 31.480 min for permethrin to 21.650 for cyfluthrin. Conclusions:The results presents here provide the status report of the insecticide resistance/ susceptibility of Cx.quinquefasciatus in major filaria endemic areas of northern India.展开更多
A comparative investigation of the resistance and ability to trigger high voltage（HV） discharge for a single filament（SF） and multiple filaments（MFs） has been carried out.The experimental results show that the t...A comparative investigation of the resistance and ability to trigger high voltage（HV） discharge for a single filament（SF） and multiple filaments（MFs） has been carried out.The experimental results show that the trend of the breakdown threshold of the SF exactly follows that of its resistance,but this is not the case for the MF.The MF＇s resistance is much smaller than the SF＇s.However,the MF shows a slightly higher HV breakdown threshold than the SF.The underlying physics is that the measured resistance of the MF is collectively contributed by every filament in the MF while the HV breakdown threshold is determined by only one single discharging path.展开更多
The aim of this study was to detect the expression of 4 clinically-important efflux pumps in the Resistance-Nodulation-Cell Division (RND) family including MexAB-OprM, MexXY, MexCD-OprJ and MexEF-OprN in Pseudomonas a...The aim of this study was to detect the expression of 4 clinically-important efflux pumps in the Resistance-Nodulation-Cell Division (RND) family including MexAB-OprM, MexXY, MexCD-OprJ and MexEF-OprN in Pseudomonas aeruginosa using a combination of resistance-phenotypic markers and multiplex RT-PCR (mRT-PCR). The antibiotic substrates specific for each Mex systems were used as phenotypic markers including carbenicillin, MexAB-OprM, erythromycin, MexCD-OprJ, norfloxacin and imipenem, MexEF-OprN and gentamicin, MexXY-OprM. The methods were validated with reference strains with known genotypes of the Mex systems and the potential applicability in clinical practice was tested with clinical isolates. The results for the reference strains support that the combination of resistance phenotype and mRT-PCR is a potential-attractive method for diagnosis of efflux-mediated resistance in P. aeruginosa. Further development to make it more practical for clinical use and study in a larger number of clinical isolates is required.展开更多
The good understanding of the mechanisms of resistance to herbicides in weeds is a necessity to implement sustainable weed management strategies. Here, a study was conducted to characterize the molecular bases of resi...The good understanding of the mechanisms of resistance to herbicides in weeds is a necessity to implement sustainable weed management strategies. Here, a study was conducted to characterize the molecular bases of resistance to acetyl coenzyme A carboxylase （ACCase） and acetolactate synthase （ALS） inhibiting herbicides in Lolium rigidum populations from Tunisia. Nine Lolium rigidum （ryegrass） populations collected in wheat fields from Northern Tunisia were investigated for their resistance to two ACCase-inhibiting herbicides and an ALS-inhibiting herbicide. All populations were tested in the greenhouse in pots using the commercial dose to determine resistance status. Survival plants were also tested for the presence of two ACCase （L 1781 and N2041） and two ALS （P197 and W574） mutant resistant alleles using molecular markers. Resistance to ACCase-inhibiting herbicides was found in all tested populations. Comparison of the results from herbicide sensitivity bioassays with genotyping indicated that more than 80% of the plants resistant to ACC-inhibiting herbicides would be resistant via increased herbicide metabolism. However, ALS-inhibiting herbicides are still more or less controlling ACCase resistant populations, so indicating that the selection process of resistance is ongoing. Target-site resistance appears to be the major mechanism for these early cases of ALS inhibitor resistance. This study reported the first case of resistance to ALS-inhibiting herbicides in ryegrass in Tunisia, and investigated the molecular bases of this resistance. It establishes the clear importance of non target-site resistance to ACCase- and/or ALS-inhibiting herbicides.展开更多
基金Supported by Agricultural Science and Technology Independent Innovation Fund of Jiangsu Province[CX(12)1003]Science and Technology Support Program of Jiangsu Province(BE2013301)Special Fund for the Construction of Modern Agriculture Industry System of China(CARS-01-47)~~
文摘Rice blast is one of the important diseases in major rice producing areas of China. The main blast resistance genes Pi-ta and Pi-b showed broad-spectrum and durable resistance to rice blast in many rice growing areas of China, which have been widely utilized in rice breeding and commercial production. In this study, on the basis of detection and verification of the genotypes of 22 rice varieties har- boring known blast resistance genes （Pi-ta and Pi-b） and blast susceptibility genes （pi-ta and pi-b）, two multiple PCR systems for these genes were established by us- ing the functional markers of blast resistance genes Pi-ta and Pi-b as well as blast susceptibility genes pi-ta and pi-b, respectively. Specifically, multiple PCR system I could simultaneously detect blast resistance genes Pi-ta and Pi-b, while system II could detect simultaneously blast susceptibility genes pi-ta and pi-b. In addition, the genotypes of 336 high generation breeding materials were detected with these two multiple PCR systems. The results were highly consistent with those of conventional single mark detection, indicating that these two multiplex PCR systems were stable, reliable and time-saving. The established multiplex PCR systems may serve as a rapid and efficient method to identify and screen rice germplasm resources and can be applied in marker-assisted selection to polymerize multiple genes for blast resis- tance in rice breeding.
基金National Natural Science Foundation of Chinese,No.3988007
文摘AIM To observe the drug sensitizing effect andrelated mechanisms of fas gene transduction onhuman drug-resistant gastric cancer cellSGC7901/VCR(resistant to Vincristine).METHODS The cell cycle alteration wasobserved by FACS.The sensitivity of gastriccancer cells to apoptosis was determined by invitro apoptosis assay.The drug sensitization ofcells to several anti-tumor drugs was observedby MTT assay.Immunochemical method wasused to show expression of P-gp and Topo Ⅱ ingastric cancer cells.RESULTS Comparing to SGC7901 and pBK-SGC7901/VCR,fas-SGC7901/VCR showeddecreasing G2 cells and increasing S cells,theG2 phase fraction of pBK-SGC7901/VCR wasabout 3.0 times that of fas-SGC7901/VCR,but Sphase fraction of fas-SGC7901/VCR was about1.9 times that of pBK-SGC7901/VCR,indicatingS phase arrest of fas-SGC7901/VCR.FACS alsosuggested apoptosis of fas-SGC7901/VCR,fas-SGC7901/VCR was more sensitive to apoptosisinducing agent VM-26 than pBK-SGC7901/VCR.MTT assay showed increased sensitization offas-SGC7901/VCR to DDP,MMC and 5-FU,butsame sensitization to VCR according to pBK-SGC7901/VCR.SGC7901,pBK-SGC7901/ VCRand fas-SGC7901/VCR had positively stainedTopo Ⅱ equally.P-gp staining in pBK- SGC7901/VCR was stronger than in SG07901,but there was little staining of P-gp in fas.SGC7901/VCR.CONCLUSION fas gene transduction couldreverse the MDR of human drug-resistant gastriccancer cell SGC7901/VCR to a degree,possiblybecause of higher sensitization to apoptosis anddecreased expression of P-gp.
文摘Multiple myeloma(MM) is a cancer caused by uncontrolled proliferation of antibody-secreting plasma cells in bone marrow, which represents the second most common hematological malignancy. MM is a highly heterogeneous disease and can be classified into a spectrum of subgroups based on their molecular and cytogenetic abnormalities. In the past decade, novel therapies, especially, the first-in-class proteasome inhibitor bortezomib, have been revolutionary for the treatment of MM patients. Despite these remarkable achievements, myeloma remains incurable with a high frequency of patients suffering from a relapse, due to drug resistance. Mutation in the proteasome β5-subunit(PSMB5) was found in a bortezomib-resistant cell line generated via long-term coculture with increasing concentrations of bortezomib in 2008, but their actual implication in drug resistance in the clinic has not been reported until recently. A recent study discovered four resistance-inducing PSMB5 mutations from a relapsed MM patient receiving prolonged bortezomib treatment. Analysis of the dynamic clonal evolution revealed that two subclones existed at the onset of disease, while the other two subclones were induced. Protein structural modeling and functional assays demonstrated that all four mutations impaired the binding of bortezomib to the 20 S proteasome, conferring different degrees of resistance. The authors further demonstrated two potential approaches to overcome drug resistance by using combination therapy for targeting proteolysis machinery independent of the 20 S proteasome.
基金supported by the National Natural Science Foundation of China [Grant agreement 31502124]the National Science and Technology Major Project of China [Grant agreement 2018ZX10733402]
文摘Objective Antimicrobial resistance(AMR)has become a global concern and is especially severe in China.To effectively and reliably provide AMR data,we developed a new high-throughput real-time PCR assay based on microfluidic dynamic technology,and screened multiple AMR genes in broiler fecal samples.Methods A high-throughput real-time PCR system with an new designed integrated fluidic circuit assay were performed AMR gene detection.A total of 273 broiler fecal samples collected from two geographically separated farms were screened AMR genes.Results The new assay with limits of detection ranging from 40.9 to 8,000 copies/reaction.The sensitivity rate,specificity rate,positive predictive value,negative predictive value and correct indices were 99.30%,98.08%,95.31%,99.79%,and 0.9755,respectively.Utilizing this assay,we demonstrate that AMR genes are widely spread,with positive detection rates ranging from 0 to 97.07%in 273 broiler fecal samples.bla CTX-M,bla TEM,mcr-1,fex A,cfr,optr A,and int I1 showed over 80%prevalence.The dissemination of AMR genes was distinct between the two farms.Conclusions We successfully established a new high-throughput real-time PCR assay applicable to AMR gene surveillance from fecal samples.The widespread existence of AMR genes detected in broiler farms highlights the current and severe problem of AMR.
基金supported by the China Agriculture Research System of MOF and MARA(CARS-12)the Open Fund of the National Key Laboratory of Crop Genetic Improvement(ZK201909)。
文摘Clubroot and herbicide resistance,high oleic acid(OA)content,and early maturity are targets of rapeseed(Brassica napus L.)breeding.The objective of this study was to develop new male-fertility restorer lines by pyramiding favorable genes to improve these traits simultaneously.Seven elite alleles for the four traits were introduced into the restorer line 621R by speed breeding with marker-assisted and phenotypic selection.Six introgression lines(ILs)were developed with four-to seven-gene combinations and crossed with two elite parents to develop hybrids.All ILs and their corresponding hybrids displayed high resistance to both clubroot pathotype 4 and sulfonylurea herbicides.Three ILs and their hybrids showed large increases in OA contents and four showed earlier maturity.These new ILs may be useful in rapeseed hybrid breeding for the target traits.
基金This work was supported by the Genetically Modified Organisms Breeding Major Project of China(2019ZX08010004–004)the National Natural Science Foundation of China(31901579).
文摘The introduction of Bacillus thuringiensis(Bt)cotton has reduced the burden of pests without harming the environment and human health.However,the efficacy of Bt cotton has decreased due to field-evolved resistance in insect pests over time.In this review,we have discussed various factors that facilitate the evolution of resistance in cotton pests.Currently,different strategies like pyramided cotton expressing two or more distinct Bt toxin genes,refuge strategy,releasing of sterile insects,and gene silencing by RNAi are being used to control insect pests.Pyramided cotton has shown resistance against different cotton pests.The multiple genes pyramiding and silencing(MGPS)approach has been proposed for the management of cotton pests.The genome information of cotton pests is necessary for the development of MGPS-based cotton.The expression cassettes against various essential genes involved in defense,detoxification,digestion,and development of cotton pests will successfully obtain favorable agronomic characters for crop protection and production.The MGPS involves the construction of transformable artificial chromosomes,that can express multiple distinct Bt toxins and RNAi to knockdown various essential target genes to control pests.The evolution of resistance in cotton pests will be delayed or blocked by the synergistic action of high dose of Bt toxins and RNAi as well as compliance of refuge requirement.
基金the funding support provided by the Department of Biotechnology(DBT),Government of India(Grant Nos.BT/AB/FG-2 (PH-II)/2009 and BT/PR11705/AGR/02/646/2008)
文摘In marker-assisted breeding for bacterial blight(BB) resistance in rice, three major resistance genes, viz., Xa21, xa13 and xa5, are routinely deployed either singly or in combinations. As efficient and functional markers are yet to be developed for xa13 and xa5, we have developed simple PCR-based functional markers for both the genes. For xa13, we designed a functional PCR-based marker, xa13-prom targeting the In Del polymorphism in the promoter of candidate gene Os8N3 located on chromosome 8 of rice. With respect to xa5, a multiplex-PCR based functional marker system, named xa5 FM, consisting of two sets of primer pairs targeting the 2-bp functional nucleotide polymorphism in the exon II of the gene TFIIA5(candidate for xa5), has been developed. Both xa13-prom and xa5 FM can differentiate the resistant and susceptible alleles for xa13 and xa5, respectively, in a co-dominant fashion. Using these two functional markers along with the already reported functional PCR-based marker for Xa21(p TA248), we designed a single-tube multiplex PCR based assay for simultaneous detection of all the three major resistance genes and demonstrated the utility of the multiplex marker system in a segregating population.
文摘Summary: The expression and functional activity of multiple drug resistance (MDR1) gene in human normal bone marrow CD34+ cells was observed. Human normal bone marrow CD34+ cells were enriched with magnetic cell sorting (MACS) system, and then liposome-mediated MDR1 gene was transferred into bone marrow CD34+ cells. Fluorescence-activated cell sorter was used to evaluate the expression and functional activity of P-glycoprotein (P-gp) encoded by MDR1 gene. It was found that the purity of bone marrow CD34+ cells was approximately (91±4.56) % and recovery rate was (72.3±2.36) % by MACS. The expression of P-gp in the transfected CD34+cells was obviously higher than that in non-transfected CD34+ cells. The amount of P-gp in non-transfected CD34+ cells was (11.2±2.2) %, but increased to (23.6±2.34) % 48 h after gene transfection (P<0.0l). The amount of P-gp was gradually decreased to the basic level one week later. The accumulation and extrusion assays showed that the overexpression of P-gp could efflux Rh-123 out of cells and there was low fluorescence within the transfected cells. The functional activity of P-gp could be inhibited by 10 μg/ml verapamil. It was suggested that the transient and highly effective expression and functional activity of P-gp could be obtained by liposome-mediated MRD1 transferring into human normal bone marrow CD34+ cells.
基金Research Projects-Joint Fund for Applied Basic Research of Kunming Medical University,Yunnan Provincial Department of Science and Technology(No.2018FE001(-113),No.202301AY070001-098)Open project of Yunnan Clinical Medical Center(Nos.2020LCZXKF-XY02,XY06,XY16+1 种基金2022LCZXKF-XY02)Yunnan Health Training Project of High Level Talents(No.D–2018018).
文摘Multiple myeloma(MM)is a hematological tumor with high mortality and recurrence rate.Carfilzomib is a new-generation proteasome inhibitor that is used as the first-line therapy for MM.However,the development of drug resistance is a pervasive obstacle to treating MM.Therefore,elucidating the drug resistance mechanisms is conducive to the formulation of novel therapeutic therapies.To elucidate the mechanisms of carfilzomib resistance,we retrieved the GSE78069 microarray dataset containing carfilzomib-resistant LP-1 MM cells and parental MM cells.Differential gene expression analyses revealed major alterations in the major histocompatibility complex(MHC)and cell adhesion molecules.The upregulation of the tumor necrosis factor(TNF)receptor superfamily member 1A(TNFRSF1A)gene was accompanied by the downregulation of MHC genes and cell adhesion molecules.Furthermore,to investigate the roles of these genes,we established a carfilzomib-resistant cell model and observed that carfilzomib resistance induced TNFRSF1A overexpression and TNFRSF1A silencing reversed carfilzomib resistance and reactivated the expression of cell adhesion molecules.Furthermore,TNFRSF1A silencing suppressed the tumorigenesis of MM cells in immunocompetent mice,indicating that TNFRSF1A may lead to carfilzomib resistance by dampening antitumor immunity.Furthermore,our results indicated that TNFRSF1A overexpression conferred carfilzomib resistance in MM cells and suppressed the expression of MHC genes and cell adhesion molecules.The suppression of MHC genes and cell adhesion molecules may impair the interaction between immune cells and cancer cells to impair antitumor immunity.Future studies are warranted to further investigate the signaling pathway underlying the regulatory role of TNFRSF1A in MM cells.
文摘Rice yellow mottle virus (RYMV) is a major biotic constraint for rice production in Africa. The resistance-breaking ability of Tanzanian RYMV strains and phylotypes (S4lm (Tz526), S4lv (Tz516), S4ug (Tz508), S5 (Tz429, Tz445), S6c (Tz486) and S6w (Tz539)) were tested by inoculating rice cultivars with RYMV1 resistant alleles (Gigante (rymv1-2), Tog12387 (rymv1-3), Tog5681 (rymv1-3), Tog5438 (rymv1-4), Tog5672 (rymv1-4+rymv2) and Tog5674 (rymv 1-5)) in a screen house. The results revealed multiple resistance-breaking strains and phylotypes on resistant cultivars Gigante, Tog12387, Tog5438 and Tog5681. However, the resistance breakdown was highly variable depending on the strain used, and disease severity ranged from 11% - 75.3%. The virulence potential of RYMV phylotype S4lm (Tz526) was similar to phylotype S6w (Tz539). The impact of strains and phylotypes on yield and its components in rice cultivars revealed highly significant differences (P ≤ 0.001). The lowest percent plant height reduction (2.8%), number of tillers per plant (2.5%), 1000 grain weight (2.7%), spikelet sterility (3.5%) and yield (5%) was recorded in rice cultivar Gigante inoculated with RYMV phylotype S6c (Tz486). Phylotype S6c (Tz486) despite being less virulent compared to other strains, its virus titer in rice cultivar Gigante (1.833) was higher than S5 (Tz429, Tz445) inoculated on Tog5674 (0.171, 0.207) and S6w (Tz539) inoculated on Tog5681 (0.283). The resistant-breaking strain S5 (Tz445) multiplied in resistant rice cultivar Tog5674 without inducing visible symptoms but showed positive reaction to ELISA with low virus titer. The strain S5 overcame wide range of resistant alleles including rymv1-2, rymv1-3, rymv1-4 and rymv1-5 resistance, with exception of rymv1-4 + rymv2. The current results gave a new perspective for future identification of resistance-breaking mutations through sequencing of the RYMV genome in infected rice cultivars and mutagenesis of an infectious viral clone useful for future RYMV resistant breeding programs.
基金The work was sup—ported by the National Natural Science Foundation of China(30371078).
文摘A multiplex PCR was developed to detect benzimidazole-resistance (BZ-R) or -susceptibility (BZ-S) in Haemonchus contortus by amplification with 4 primers of a sequence of the GRU-1 gene of β-tubulin of H. contortus making use of sequence information available in Genbank. The method was based on two allele-non-specific primers and two allele- specific primers. F1 (264 bp) and F3 (799 bp) should be produced in BZ-R, F2 (585 bp) and F3 in BZ-S. With this method, we demonstrated that H. contortus BZ-R strain from Australia showed F1 and F3, and the worm BZ-S strain from Shanghai did F2 and F3. Sequence analysis of the isotype 1 gene of β-tubulin of BZ-R from Australia and BZ-S from Shanghai showed the code in residue 200 of the gene was respectively TAC and TTC. The LD50 of albendazole of the Australian BZ- R strain was 0.54 μg mL-1, the Shanghai BZ-S strain was only 0.0023 μg mL-1 by EHA (egg hatch assay). The multiplex PCR could determinate the genotype of single adult worm or several third stage larvae and was performed on at least 50 ng of genomic DNA. BZ-R H. contortus were not detected in Shihezi and Yining of the Xinjiang, Wuhe of the Anhui Province, Nanjing and Xuzhou of the Jiangsu Province. The LD50 of the H. contortus from these locations to albendazole as determined by EHA varied between 0.0023-0.0032 μg mL-1. The result indicated that the multiplex PCR could be used to differentiate BZ-R and BZ-S of H. contortus and that the BZ-R situation of H. contortus was not serious in China.
基金supported by laboratory startup funds from Peking University and the Center for Life Sciences,as well as the grants from the National Science Foundation of China(32302455 and 32270672)。
文摘CRISPR homing gene drives have considerable potential for managing populations of medically and agriculturally significant insects.They operate by Cas9 cleavage followed by homology-directed repair,copying the drive allele to the wild-type chromosome and thus increasing in frequency and spreading throughout a population.However,resistance alleles formed by end-joining repair pose a significant obstacle.To address this,we create a homing drive targeting the essential hairy gene in Drosophila melanogaster.Nonfunctional resistance alleles are recessive lethal,while drive carriers have a recoded“rescue”version of hairy.The drive inheritance rate is moderate,and multigenerational cage studies show drive spread to 96%–97%of the population.However,the drive does not reach 100%due to the formation of functional resistance alleles despite using four gRNAs.These alleles have a large deletion but likely utilize an alternate start codon.Thus,revised designs targeting more essential regions of a gene may be necessary to avoid such functional resistance.Replacement of the rescue element’s native 3'UTR with a homolog from another species increases drive inheritance by 13%–24%.This was possibly because of reduced homology between the rescue element and surrounding genomic DNA,which could also be an important design consideration for rescue gene drives.
基金supported by the National Natural Science Foundation of China (51874281)the Graduate Innovation Program of China University of Mining and Technology (2022WLKXJ006)the Postgraduate Research&Practice Innovation Program of Jiangsu Province (KYCX22_2612).
文摘In the present research,we proposed a scheme to address the issues of severe heat damage,high energy consumption,low cooling system efficiency,and wastage of cold capacity in mines.To elucidate the seasonal variations of environmental temperature through field measurements,we selected a high-temperature working face in a deep mine as our engineering background.To enhance the heat damage control cability of the working face and minimize unnecessary cooling capac-ity loss,we introduced the multi-dimensional heat hazard prevention and control method called"Heat source barrier and cooling equipment".First,we utilize shotcrete and liquid nitrogen injection to eliminate the heat source and implemented pressure equalization ventilation to disrupt the heat transfer path,thereby creating a heat barrier.Second,we establish divi-sional prediction models for airflow temperature based on the variation patterns obtained through numerical simulation.Third,we devise the location and dynamic control strategy for the cooling equipment based on the prediction models.The results of field application show that the heat resistance and cooling linkage method comply with the safety requirement throughout the entire mining cycle while effectively reducing energy consumption.The ambient temperature is maintained below 30℃,resulting in the energy saving of 10%during the high-temperature period and over 50%during the low-temperature period.These findings serve as a valuable reference for managing heat damage in high-temperature working faces.
基金The research was supported by the Hainan Provincial Joint Project of Sanya Yazhou Bay Science and Technology City(No.2021CXLH0009)the National Natural Science Foundation of China(Nos.41976222 and 42006195)+4 种基金the Scientific Research Foundation of Hainan Tropical Ocean University(No.RHDRC202201)the Science and Technology Project of Yazhou Bay Innovation Institute of Hainan Tropical Ocean University(No.2022CXYZD002)the National Key Research and Development Program of China(No.2019YFD0901104)the Open Fund for the Key Laboratory of Global Change and Marine-Atmospheric Chemistry(No.GCMAC 2010)the Open Fund of State Key Laboratory of Environmental Chemistry and Ecotoxicology(No.KF2018-05).
文摘Coastal lagoons provide many important services to human society,but their year-round use for aquaculture introduces large amounts of sewage.The contamination of antibiotic resistance genes(ARGs)is therefore of great concern.In this study,50 ARGs subtypes,two integrase genes(intl1,intl2),and 16S rRNA genes were detected by high-throughput quantitative PCR,and standard curves of all target genes were prepared for quantification.The occurrence and distribution of ARGs in a typical coastal lagoon(XinCun lagoon,China)were comprehensively explored.We detected 44 and 38 subtypes of ARGs in the water and sediment,respectively,and discuss the various factors influencing the fate of ARGs in the coastal lagoon.Macrolides-lincosamides-streptogramins B was the primary ARG type,and macB was the predominant subtype.Antibiotic efflux and antibiotic inactivation were the main ARG resistance mechanisms.The XinCun lagoon was divided into eight functional zones.The ARGs showed a distinct spatial distribution owing to the influence of microbial biomass and anthropogenic activity in different functional zones.Fishing rafts,abandoned fish ponds,the town sewage zone,and mangrove wetlands provided a large quantity of ARGs to the XinCun lagoon.Nutrients and heavy metals also significantly correlated with the fate of the ARGs,especially NO_(2)^(−)-N and Cu,which cannot be ignored.It is noteworthy that lagoon-barrier systems coupled with persistent pollutant inputs result in coastal lagoons acting as a“buffer pool”for ARGs,which can then accumulate and threaten the offshore environment.
文摘Drug resistance is becoming a great problem in developing countries due to excessive use and misuse of antibiotics. The emergence of new pathogenic strains with resistance developed against most of the antibiotics which may cause,difficult to treat infection.To understand the current scenario in different mode of infection is most important for the clinicians and medical practitioners.This article summarized some common infections and antibiotic resistance pattern found among these pathogens.
基金supported by the internal funds of Biotechnology Department,AMU,Aligarh
文摘This study was to investigate the current trends of multiple drug resistance in bacteria against antibiotics for the proper empirical treatmen.Clinical isolates were collected from community-acquired infection of urinary tract patients in Aligarh India from March 1999 to August 1999.Antibiotic susceptibility test was performed,using the disc diffusion method followed by plasmid isolation by the method of Kado and Liu.Transfer experiments were performed by the method of Lederberg and Cohen.Clinical study revealed that this infection was more common in young women.Various strains of E.coli isolated during the course of study were found to show multiple antibiotic resistance which was further characterized as plasmid-borne drug resistance.This study shows that E.coli may be one of the important causative agents of urinary tract infection(UTI) in young women.
文摘Objective:To understand the insecticide resistance status of Culex quinquefasciatus Say（Diptera: Culicidae）（Cx.Quinquefasciatus） to deltamethrin,cyfluthrin,permethrin,lambdacyhalothrin, DDT and malathion in filarial endemic areas of Uttar Pradesh,India.Methods:Insecticide susceptibility assays were performed on wild-caught adult female Cx.quinquefasciatus mosquitoes to deltamethrin（0.05%）,cyfluthrin（0.15%）,permethrin（0.75%）,lambdacyhalothrin （0.05%）,malathion（5.0%） and DDT（4.0%）,the discriminating doses recommended by the World Health Organisation（WHO）.Results:The data showed that Cx.quinquefasciatus is highly resistant to DDT and malathion;the mortality was 28.33%and 27.5%,respectively and incipient resistance to synthetic pyrethroids（deltamethrin,cyfluthrin,permethrin,and lambdacyhalothrin）, where mortality ranged from 95.83%in permethrin to 98.33%in cyfluthrin and lambdacyhalothrin. Knockdown times(KDT<sub>50</sub>) in response to synthetic pyrethroids varied significantly between different insecticides（P【0.01） from 31.480 min for permethrin to 21.650 for cyfluthrin. Conclusions:The results presents here provide the status report of the insecticide resistance/ susceptibility of Cx.quinquefasciatus in major filaria endemic areas of northern India.
基金Project supported by the National Natural Science Foundation of China (Grant Nos. 11074027,60978014,61178022,11274053,and 11211120156)the Funds from Sci. & Tech. Deparment of Jilin Province,China (Grant No. 20111812)
文摘A comparative investigation of the resistance and ability to trigger high voltage（HV） discharge for a single filament（SF） and multiple filaments（MFs） has been carried out.The experimental results show that the trend of the breakdown threshold of the SF exactly follows that of its resistance,but this is not the case for the MF.The MF＇s resistance is much smaller than the SF＇s.However,the MF shows a slightly higher HV breakdown threshold than the SF.The underlying physics is that the measured resistance of the MF is collectively contributed by every filament in the MF while the HV breakdown threshold is determined by only one single discharging path.
文摘The aim of this study was to detect the expression of 4 clinically-important efflux pumps in the Resistance-Nodulation-Cell Division (RND) family including MexAB-OprM, MexXY, MexCD-OprJ and MexEF-OprN in Pseudomonas aeruginosa using a combination of resistance-phenotypic markers and multiplex RT-PCR (mRT-PCR). The antibiotic substrates specific for each Mex systems were used as phenotypic markers including carbenicillin, MexAB-OprM, erythromycin, MexCD-OprJ, norfloxacin and imipenem, MexEF-OprN and gentamicin, MexXY-OprM. The methods were validated with reference strains with known genotypes of the Mex systems and the potential applicability in clinical practice was tested with clinical isolates. The results for the reference strains support that the combination of resistance phenotype and mRT-PCR is a potential-attractive method for diagnosis of efflux-mediated resistance in P. aeruginosa. Further development to make it more practical for clinical use and study in a larger number of clinical isolates is required.
文摘The good understanding of the mechanisms of resistance to herbicides in weeds is a necessity to implement sustainable weed management strategies. Here, a study was conducted to characterize the molecular bases of resistance to acetyl coenzyme A carboxylase （ACCase） and acetolactate synthase （ALS） inhibiting herbicides in Lolium rigidum populations from Tunisia. Nine Lolium rigidum （ryegrass） populations collected in wheat fields from Northern Tunisia were investigated for their resistance to two ACCase-inhibiting herbicides and an ALS-inhibiting herbicide. All populations were tested in the greenhouse in pots using the commercial dose to determine resistance status. Survival plants were also tested for the presence of two ACCase （L 1781 and N2041） and two ALS （P197 and W574） mutant resistant alleles using molecular markers. Resistance to ACCase-inhibiting herbicides was found in all tested populations. Comparison of the results from herbicide sensitivity bioassays with genotyping indicated that more than 80% of the plants resistant to ACC-inhibiting herbicides would be resistant via increased herbicide metabolism. However, ALS-inhibiting herbicides are still more or less controlling ACCase resistant populations, so indicating that the selection process of resistance is ongoing. Target-site resistance appears to be the major mechanism for these early cases of ALS inhibitor resistance. This study reported the first case of resistance to ALS-inhibiting herbicides in ryegrass in Tunisia, and investigated the molecular bases of this resistance. It establishes the clear importance of non target-site resistance to ACCase- and/or ALS-inhibiting herbicides.
