A quantitative RT-PCR assay for rapid detection of Eurasianlineage H10 subtype influenza A virus

Dear Editor,Influenza A viruses(IAVs)are single-stranded,negative sense RNA viruses.IAV subtype is determined on the basis of the viral surface glycoproteins,hemagglutinin(HA),and neuraminidase(NA).To date,18 HA and 11NA subtypes have been reported(Tong et al.,2012).

Effect of electroacupuncture on the mRNA and protein expression of Rho-A and Rho-associated kinase Ⅱ in spinal cord injury rats

Electroacupuncture is beneficial for the recovery of spinal cord injury, but the underlying mechanism is unclear. The Rho/Rho-associated kinase（ROCK） signaling pathway regulates the actin cytoskeleton by controlling the adhesive and migratory behaviors of cells that could inhibit neurite regrowth after neural injury and consequently hinder the recovery from spinal cord injury. Therefore, we hypothesized electroacupuncture could affect the Rho/ROCK signaling pathway to promote the recovery of spinal cord injury. In our experiments, the spinal cord injury in adult Sprague-Dawley rats was caused by an impact device. Those rats were subjected to electroacupuncture at Yaoyangguan（GV3）, Dazhui（GV14）, Zusanli（ST36） and Ciliao（BL32） and/or monosialoganglioside treatment. Behavioral scores revealed that the hindlimb motor functions improved with those treatments. Real-time quantitative polymerase chain reaction, fluorescence in situ hybridization and western blot assay showed that electroacupuncture suppressed the m RNA and protein expression of Rho-A and Rho-associated kinase Ⅱ（ROCKⅡ） of injured spinal cord. Although monosialoganglioside promoted the recovery of hindlimb motor function, monosialoganglioside did not affect the expression of Rho-A and ROCKⅡ. However, electroacupuncture combined with monosialoganglioside did not further improve the motor function or suppress the expression of Rho-A and ROCKⅡ. Our data suggested that the electroacupuncture could specifically inhibit the activation of the Rho/ROCK signaling pathway thus partially contributing to the repair of injured spinal cord. Monosialoganglioside could promote the motor function but did not suppress expression of Rho A and ROCKⅡ. There was no synergistic effect of electroacupuncture combined with monosialoganglioside.

Population Dynamics and Survival of Rhizoctonia solani AG-1 in Field Soil Under Rice-Wheat Rotation

A field under rice-wheat rotation was selected near Chengdu, China, to study thepopulation of Rhizoctonia solani anastomosis group 1 (AG-1), pathogen causing ricesheath blight disease, in natural soil ecosystem. Inocula of the fungus recovered fromthe field were divided into three types, i.e., sclerotia, free mycelium retained in thesoil passed through a 0.355mm sieve, and colonized plant debris which was subdividedinto small colonized debris retained between 2.00 and 0.355mm sieves and large colonizeddebris retained on 2.00mm sieve after wet screening. Quantitative estimation of thethree types of inocula in one year indicated that small colonized debris was the dominantinoculum type for most of the time. The population peaked in March and September at 1210and 480 colonized debris 100g-1 air-dry soil respectively, and fell down in December andAugust to 0 and 177 colonized debris 100g-1 air-dry soil respectively. Free mycelium wasonly detectable in March, September and October with 1209, 7.9 and 14.5g fresh wtmyceliumg-1 air-dry soil respectively, which corresponded to the two peaks and the secondhighest level of small debris density in the year. Viable sclerotia and large colonizeddebris were rare with populations ranging from 0 to 3 for sclerotia and 0 to 14 for largecolonized debris 100g-1 air-dry soil, but were the main structures to survive overwinter. It was expected that soil temperature was the main factor determining populationdynamics of R.solani AG-1 in natural soil. Optimum temperature for population increasingis predicted to be around 15℃, with a range from 10 to 25℃. Viability tests indicatedthat 60.9% sclerotia could survive after 265d being buried in natural sandy loam in fieldconditions in Beijing, while colonized rice straw debris (0.5-1.0cm long) could notyield the fungus on medium plates after 88d of being buried under the same conditions.

An analytical method for quantitative estimation of the cytotoxicity of dental alloys using MTT assay,ICP analysis and effective cytotoxicity calculation

The two most important criteria for dental materials are their biofunctional and biocompatible endurance within the anticipated life-span of the dental restoration in the mouth. Biocompatibility relates mainly to the allergenicity and the toxicity of the material. To test the non-specific toxicity of dental materials, in vitro cell culture assays have been developed. For in vitro screening, such tests are recommended to check the cytotoxicity of dental materials (ISO 10993 5). Various studies have already been performed to quantitatively determine the cytotoxicity level of dental alloys. However, as long as only dental alloys and the cell culture technique are applied, it is not possible to determine which of the alloying elements cause the cytotoxicity. Therefore, an analytical method is needed. Wataha et al determined in 1991 the TC50 values of 9 metal cations of various dental casting alloys, using cell culture methods. Kapert et al reported in 1994 a complex in vitro test concept, where the ICP analysis (inductively coupled plasma emission spectroscopy) was introduced to measure the trace elements extracted from various alloys. Experimentelle Zahnheilkunde, Universitts ZMK Klinik Freiburg, Germany (Lü XY and Kappert HF) The aim of the present study was to find a relation between the ICP results, the TC50 value of metal cations, and the cytotoxicity of dental alloys. The cytotoxicity levels of various dental alloys and the TC50 values of 10 metal cations were established using the MTT assay, an effective cell culture of method. Then, the concentrations of the corrosively soluted metal cations in the extracts of the alloys were measured using the ICP method. From all these experimental results it was found that the relation between the effective cytotoxicity Z eff of an alloy, the concentrations C i of i th trace element and the TC50 values T Ci of the i th metal cation can approximately be expressed by Z eff =∑iC i2·T Ci . Two significant applications of this expression are a) The cytotoxicity of an alloy can be estimated by ICP analysis of the extract if the TC50 values of the trace elements are know. b) The cytotoxicity of a new-developed-alloy can be estimated in advance, according to the alloying components.

Immune fluorescence test strips based on quantum dots for rapid and quantitative detection of carcino-embryonic antigen

At present,many researchers focused on the point-of-care testing（POCT）,a method of disease markers detection without large-scale instruments and specialized persons.However,most POCT diagnostic methods were suffered from poor detection sensitivity or inefficiency in quantitative detection.Herein,we developed a newly QD-immune fluorescence test strips（QD-IFTS） based on quantum dots（QDs） as the fluorescence nanocarrier to prepare the immune fluorescence probes in the classical immunochromatography detection system for sensing carcino-embryonic antigen（CEA）,a kind of glycoprotein produced by intestinal tissue and a broad spectrum of tumor marker for cancer diagnosis.And we designed a homemade strips fluorescence reader for detection of fluorescence intensity of QDs on the QD-IFTS.Under the optimized reaction conditions,chromatographic time of the newly QD-IFTS was only25 min,sample volume of the newly QD-IFTS was only 40 m L and the LOD of the newly QD-IFTS was 0.72 ng/m L.In addition,the efficiency and robustness of the newly QD-IFTS were confirmed by successfully application in 300 clinical serum samples,and the results revealed great potential in clinical POCT of other biomarkers.

AuNP aggregation-induced quantitative colorimetric aptasensing of sulfadimethoxine with a smartphone

A gold nanoparticle(AuNP)aggregation-induced colorimetric aptasensing method for quantitative detection of sulfadimethoxine(SDM)with a smartphone was developed.AuNPs were complexed with aptamers which protected Au NPs from aggregating in high-concentration salt solutions.In the presence of SDM,SDM bound with the aptamer on the surface of Au NPs with higher affinity,which competitively desorbed the aptamer from the AuNP surface and resulted in AuNPs aggregation,accompanied with a color change from red to purple-blue.The R,G and B values of images taken by a smartphone camera were analyzed with an app on the smartphone,and were utilized for quantitative analysis of SDM.Under the optimized conditions,the colorimetric aptasensing method using a smartphone showed high sensitivity for SDM,with the limit of detection of 0.023 ppm,lower than the allowed maximum SDM residue limit.This study provides a simple,fast,and easy to read method for on-site quantitative biochemical and cellular analysis.