Primer Exchange Reaction (PER)-Based Construction of Scaffold for Low-Speed Centrifugation-Based Isolation and Quantitative Analysis of P. aeruginosa and its application in analyzing uterine secretions with intrauterine adhesion.

Efficient isolation and sensitive quantification of Pseudomonas aeruginosa (P. aeruginosa) are crucial for identifying intrauterine infections and preventing the occurrence of intrauterine adhesion (IUA). However, traditional approaches, such as culture-based approach, are time-consuming. Herein, we constructed a detection scaffold by using primer exchange reaction (PER) that integrated the low-speed centrifugation-based isolation and sensitive quantification of target pathogenic bacteria. The established approach possesses several advantages, including (i) the approach is capable of simultaneous isolation and sensitive quantification of target bacteria; (ii) low-speed centrifugation or even manual equipment could be used to isolate target bacteria; and (iii) a low limit of detection was obtained as 54 cfu/mL. Based on this, the approach is a promising approach in analyzing P. aeruginosa from uterine secretions with IUA.

An efficient strategy for establishing a model of sensorineural deafness in rats.

Ototoxic drugs can be used to produce a loss of cochlear hair cells to create animal models of deafness. However, to the best of our knowledge, there is no report on the establishment of a rat deafness model through the combined application of aminoglycosides and loop diuretics. The aim of this study was to use single or combined administration of furosemide and kanamycin sulfate to establish rat models of deafness. The rats received intravenous injections of different doses of furosemide and/or intramuscular injections of kanamycin sulfate. The auditory brainstem response was measured to determine the hearing threshold after drug application. Immunocytochemistry and confocal microscopy were performed to evaluate inner ear morphology. In the group receiving combined administration of furosemide and kanamycin, the auditory brainstem response threshold showed significant elevation 3 days after administration, higher than that produced by furosemide or kanamycin alone. The hair cells showed varying degrees of injury, from the apical turn to the basal turn of the cochlea and from the outer hair cells to the inner hair cells. The spiral ganglion cells maintained a normal morphology during the first week after the hair cells completely disappeared, and then gradually degenerated. After 2 months, the majority of spiral ganglion cells disappeared, but a few remained. These findings demonstrate that the combined administration of furosemide and kanamycin has a synergistic ototoxic effect, and that these drugs can produce hair cell loss and hearing loss in rats. These findings suggest that even in patients with severe deafness, electronic cochlear implants may partially restore hearing.

A rapid and sensitive method for the detection of aromatic amines in cosmetics.

Aromatic amines (AAs) are common chemical pollutants and banned ingredients in cosmetics. In this study, a rapid, simple and stable method for the detection of nine AAs in cosmetics was established based on the optimization of cation exchange solid-phase extraction and liquid chromatography tandem mass spectrometry. The method displayed good linearity within a range of 2-1,000 µg/kg, with limits of quantitation at the level of µg/kg for cosmetic samples. The recoveries obtained for all analyzed amines ranged between 83.6 and 97.8%, and the repeatability (r) and reproducibility (R) values indicated that all nine AAs showed good precision (r ≤ 4.5% and R ≤ 7.7%). The method was applied for the detection of 36 cosmetic samples. It was found that the primary pollutants of AAs were 3, 3'-dichlorobenzidine and 4-aminoazobenzene. The total amine concentration in cosmetic samples ranged from 880 to 5,200 µg/kg. The proposed method is applicable for the analysis of most cosmetic samples.

[Serological characteristics and transfusion efficacy evaluation in 61 cases of autoimmune hemolytic anemia].

This study was aimed to analyze the serological characteristics, efficacy and safety of incompatible RBC transfusion in patients with autoimmune hemolytic anemia (AIHA). The patients with idiopathic or secondary AIHA were analyzed retrospectively, then the serological characteristics and the incidence of adverse transfusion reactions were investigated, and the efficacy and safety of incompatible RBC transfusion were evaluated according to the different autoantibody type and infused different RBC components. The results showed that out of 61 cases of AIHA, 21 cases were idiopathic, and 40 cases were secondary. 8 cases (13.1%) had IgM cold autoantibody, 50 cases (82.0%) had IgG warm autoantibody, and 3 cases (4.9%) had IgM and IgG autoantibodies simultaneously. There were 18 cases (29.5%) combined with alloantibodies. After the exclusion of alloantibodies interference, 113 incompatible RBC transfusions were performed for 36 patients with AIHA, total efficiency rate, total partial efficiency rate and total inefficiency rate were 56.6%, 15.1% and 28.3%, respectively. Incompatible RBC transfusions were divided into non-washed RBC group and washed RBC group. The efficiency rate, partial efficiency rate and inefficiency rate in non-washed RBC group were 57.6%, 13.0% and 29.4%, respectively. The efficiency rate, partial efficiency rate and inefficiency rate in washed RBC group were 53.6%, 21.4% and 25.0%, respectively. There was no significant difference of transfusion efficacy (P > 0.05) in two groups. Incompatible RBC transfusions were also divided into IgM cold autoantibody group and IgG warm autoantibody group. The efficiency rate, partial efficiency rate and inefficiency rate in IgM cold autoantibody group were 46.2%, 30.8% and 29.4%, respectively. The efficiency rate, partial efficiency rate and inefficiency rate in IgG warm autoantibody group were 56.7%, 13.4% and 29.9%, respectively. There was no significant difference of transfusion efficacy (P > 0.05 ) in two groups. Hemolytic transfusion reaction was not observed in all incompatible RBC transfusions. It is concluded that the same ABO type of non-washed RBC transfusion and O type washed RBC transfusion are all relatively safe for the AIHA patients with severe anemia after the exclusion of alloantibodies interference. There is no significant difference of transfusion efficacy in two groups. The same ABO type of non-washed RBC transfusion is more convenient and efficient than washed RBC transfusion, and excessive use of type O RBCs can also be avoided.

[Establishment of genotyping method for fetal ABO group from pregnant maternal peripheral blood].

This study was aimed to establish a genotyping method to detect ABO group gene of fetus from peripheral blood of pregnant women for prenatal diagnosis of hemolytic disease of newborn (HDN) resulting from ABO blood group incompatibility. 4 pairs of primers were designed according to ABO blood group gene DNA and mRNA sequences. 20 plasma DNA samples from healthy donors were extracted and amplified to explore the best conditions for plasma DNA extraction and PCR amplification. The O group plasma DNA was mixed with A group or B group plasmas by the ratios of 1:1, 2:1, 4:1, 8:1, 10:1, 20:1, 40:1, 100:1 to simulate the status of mixed ABO gene from pregnant maternal blood and to establish the mixed blood group ABO genotyping technology. The pregnant maternal blood samples with more than 30 weeks of gestation were selected for detecting the fetal ABO blood group genotype. The blood samples should be taken as possible as after birth for identification of ABO blood group and evaluation of sensitivity and accuracy of fetal ABO blood group genotyping technology through peripheral blood of pregnant women. The results indicated that the minimal amount of template DNA from single blood plasma for accuracy identification was at least about 0.625 ng, the DNA amount extracted from 500 microl of plasma could meet the requirement for PCR amplification. When the proportion of O group plasma DNA in mixed plasma DNA was

[Application of thrombelastography in evaluation of platelet function during storage].

This study was aimed to explore changes of platelet function in vitro during storage by thrombelastography (TEG). 12 units plateletpheresis were randomly selected and stored at 20 to 24 degrees C with agitation. Thrombelastography variable parameters R, K values and maximal amplitude (MA) were measured on 1, 2, 3, 4, 5 days of platelet storage. Platelet concentration, mean platelet volume (MPV), hypotonic shock response (HSR), CD62p expression and CD62p reexpression on platelet surface were detected at the same time. Changes of platelet function in virto were systematically evaluated by above-mentioned indexes. The results showed that MPV augmented slightly with prolongation of preserved time (p > 0.05), and CD62p expression on platelet surface increased remarkably (p < 0.01), while CD62p reexpression decreased gradually (p < 0.01). There were no significant differences in HSR level of platelets during storage (p > 0.05). R value increased with prolongation of preserved time (p < 0.01). There were no obvious changes on K value and alpha Angle during storage (p > 0.05). There were no obvious changes in MA from 1 to 4 days, and MA decreased slightly on day 5 (p < 0.05). It is concluded that there was no significant change in MA and HSR which reflects comprehensive coagulation of platelets during storage. Platelets on the end of storage have excellent function of hemostasis; Thrombelastography parameter MA value can be used as a valuable indicator for evaluation of platelet function in vitro during storage.