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Arch BiochemMol Biol2019;10(4):035-051DOI: 10.26502/abmb.007Archives of Biochemistry and Molecular BiologyVol. 10 No. 4 -December2019. 36AbstractArgonaute2 (AGO2) is a core catalytic component of the RNA-induces silencing complex (RISC) that binds to small guide RNAs containing small interfering RNA (siRNA) and microRNA (miRNA). The guide RNA leads RISC to the complementary mRNA for gene suppression. We cloned the full length cDNA (2193 bp) of the Ago2gene (PxAgo2) from diamondback moth (DBM, Plutella xylostella). The predicted PxAgo2 protein hadan83 kDa molecular weight withatheoretical isoelectric point of 9.39. The phylogenetic tree showed a high similarity of PxAgo2with Bombyx moriAgo2 (BmAgo2). Western blot and RT-qPCR analyses showed a clear increase in the PxAgo2 mRNA and protein expression levels in the egg, 4thinstar larva, pre-pupa, pupa and adult. The double-stranded RNA-mediated RNAi of PxAgo2in DBM larvae was found 3 h after dsRNA injection,and the knockdown level was increased over time up to 36 h. PxAgo2silencing recovered the expression of PxBurs-?(Bursicon-? inDBM) to the normal expression level, which was suppressed bydsPxBurs-?in a DBM cell line. The overexpression of PxAgo2fundamentally enhanced the PxBurs-? silencing efficiency in DBM cells. Our findings reveal that PxAgo2is involved in the dsRNA-regulated gene silencing mechanism and performs a crucial function in the RNAi process ofDBM
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Objective To realize hand hygiene status and effect of hand-drying measures on hand-washing of health care workers(HCWs)in an intensive care unit(ICU).Methods From February to April 2013,210 HCWs in an ICU were selected and randomly divided into three groups,group A dried hands with paper towel,group B with hand drier,and group C with personal towel,specimens from hands before hand-washing,after hand-washing,and after hand-drying were taken and performed detection.Results Hand microorganism count in group A,B and C before washing hands after contacting patients was (29.10±15.08)CFU/cm2 ,(31.42±14.76)CFU/cm2 and (30.36±15.52) CFU/cm2 respectively,the difference was not statistically different(F=0.048,P >0.05);After six-step hand-washing, hand microorganism count before hand drying in group A,B and C was (3.26 ±0.61 )CFU/cm2 ,(2.98 ±0.59) CFU/cm2 and (3.87±0.67)CFU/cm2 respectively,compared with hand microorganism count before hand-washing, the difference was statistically different(all P <0.01 ).After adopting different hand-drying methods,microorgan-ism count in three groups was statistically different(F =1 .892,P <0.05),group A ([1 .29±0.58]CFU/cm2 )was significantly lower than group B and C,group B ([9.51 ±0.73 ]CFU/cm2 )was significantly lower than group C ([22.76±4.11]CFU/cm2 );the qualified rate in group A (90.00%)was significantly higher than group B and C, group B (68.57%)was significantly higher than group C (47.14%).The top 5 pathogens isolated from HCWs’ hands were Pseudomonas aeruginosa ,Acinetobacter baumannii ,Klebsiella pneumoniae ,Escherichia coli ,and Coag-ulase negative Staphylococcus ,these strains were highly consistent with the top 5 multidrug-resistant organisms (MDROs)isolated from ICU patients in this hospital in 2013.Conclusion HCWs’hands are seriously contamina-ted after all kinds of medical performance;if hands are improperly dried,secondary contamination may occur;patho-gens isolated from HCWs’hands are highly consistent with MOROs from patients,timely and correct hand-washing and hand-drying is the key link to ensure the quality of hand hygiene,and is of great significance to reduce the occur-rence of MDROs infection in ICU patients.
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By establishing experimental modal of laryngeal allograft,the short-term histopathological changes of liver and kidney in Cyclosporine A (CsA)-treated rats receiving laryn- geal allograft were observed.The animals were divided into 3 groups.Group 1 was given CsA 15 mg?kg~(-1)/d by daily intraperitoneal injection for 2 weeks,Group 2 received CsA 25 mg?kg~(-1)/d, and the third group without CsA treatment served as control.All of recipients were sacrificed 14 days after transplantation.Histological examination showed that CsA nephrotoxicity was charac- terized by abundant vacuolation of the proximal tubular epithelium cells,hyaline regeneration of arterioles with thickening of vascular wall,and striped interstitial fibrosis and its hepatotoxicity by fatty degeneration with mild hyperplasia of Kupffer's cells and focal necrosis of hepatocytes. Histopathological changes of CsA-induced hepato- and nephrotoxicity of the recipients were closely correlated to the dosage of CsA received.
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A rat model of laryngeal allotransplantation was established by employing an end- to-side anastomosis of donor bilateral common carotid arteries to recipient common carotid artery and external jugular vein separatively.Thirteen allotransplants were performed in 26 SD rats.All recipients survived.Three days after the operation,a visible fibrinous adherence was observed around the laryngeal grafts.By the 7th day,the adherence became intensified.The airway was plugged with mucoid material,and the viable grafts were surrounded by connective tissues.Our results confirmed that the rat model was practicable for laryngeal allograft.Besides the difference of the major histocompatibility between the donor and recipient,the skill of microsurgery,the prevention of infections and the methods of donor organ flushing are all vital to a successful trans- plantation.
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ve To establish a method of high performance liquid chromatography (HPLC) for determi-nation of nitrite in water. Methods The trace content of nitrite in water sample was determined by indirect UV-HPLC. The mixture of methanol/o-phthalic acid (pH value was adjusted to 8.6 by 0.10 mol/L NaOH solution)5: 95 was defined as mobile phase. The water sample was directly filtered by chromatographic column pack with ODP. Results Under the conditions of wave length of 270 nm and flow rate of 0.9 ml/ min, the linear range of this assay was 0.0~20.0?g/ml nitrite, the relative standard deviation was 3.9%. The average recovery rate and detection limit were 99.9% and 0.001?g/ ml respectively. Conclusion This method could be applied to the determination of trace amount of nitrite in drinking water, purified water and mineral water.