Interference of CD38 monoclonal antibody in blood compatibility testing and its countermeasures: A general consensus among experts / 中国输血杂志

With continuous discovery of tumor immune targets and continuous changes in antibody research and development technology, antibody drugs are becoming more and more widely used in clinical practice. However, some targets are not only expressed on tumor cells, but also on red blood cells. Therefore, the clinical application of antibodies against the corresponding targets may interfere with the detection of blood transfusion compatibility, resulting in difficulty in blood matching or delay of blood transfusion. This consensus summarizes the current solutions for the interference of CD38 monoclonal antibody (CD38 mAb) in transfusion compatibility testing. After analyzing the advantages and disadvantages of different methods, polybrene and sulfhydryl reducing agents [dithiothreitol (DTT) or 2-mercaptoethanol (2-Me)], as a solution for CD38 mAb interference in blood compatibility testing, are recommended for Chinese patients, so as to eliminate blood transfusion interference produce by CD38 mAb and further provide a pre-transfusion workflow for clinicians and technicians in Department of Blood Transfusion.

Evaluation of dot-blot test for serological diagnosis of bovine brucellosis

Abstract The objective of this study was to standardize and validate the dot-blot test for the serological diagnosis of bovine brucellosis, compare the results with those found in the 2-mercaptoethanol (2-ME) and complement fixation test (CF), and estimate the relative sensitivity and specificity of the dot-blot compared to these tests. Fifty bovine blood serum samples were used for the test standardization, and 1315 samples were used for evaluation and comparison between the tests; the results were compared using the Kappa indicator. At the end of standardization, it was established as optimal for the antigen obtained from Brucella abortus B19 after passing through a microorganism rupture process, the blood serum samples diluted at 1:100, and the conjugate at 1:30,000. The comparison of the dot-blot results with 2-ME showed Kappa index of 0.9939, sensitivity of 99.48%, and specificity 99.91%, with CF, Kappa index of 0.8226, sensitivity 100% and specificity 95.32%. Using the combination of the test results 2-ME and CF to establish the true condition of the animal, the dot-blot showed relative sensitivity of 100%, and relative specificity of 99.91%. The evaluated test proved to be effective and reliable, besides being easy to handle and interpret the results.

Protective Effects of Hydroxysafflor Yellow A against Oxidative Damage of β-Mercaptoethanol During Neural Differentiation of Mesenchymal Stem Cells / 中草药·英文版

Objective To study the protective effects of hydroxysafflor yellow A (HSYA) against the oxidative damage caused by β-mercaptoethanol (BME) during neural differentiation of mesenchymal stem cells (MSCs) in vitro. Methods When the confluence reached 50%-60%, 4th passage MSCs were divided into three groups to culture. G1: normal group which was cultured using basic medium (DMEM containing 10% FBS all the time); G2: unprotected group which was continuously cultured using basic medium for 24 h, and then cultured using pre-induction medium (DMEM containing 10% FBS and 1 mmol/L BME); G3: protected group which was firstly cultured using protective medium (DMEM containing 10% FBS and 160 mg/L HSYA) for 24 h, and then cultured using pre-induction medium for 24 h. After these treatments as above, cell viability, relative levels of SOD/GSH and apoptosis rate were respectively detected. The expression of Bcl and Bax was examined by Western blotting. After HSYA protection and BME pre-induction, neural induction was performed. The expression of NSE and MAP-2 was respectively analyzed on cellular and molecular levels. Results Compared with unprotected group, 160 mg/L HSYA could obviously improve cells viability, maintain high level of SOD and GSH in MSCs, reduce apoptosis rate and improve the ratio of Bcl/Bax. After protection with 160 mg/L HSYA, the survival time of neuron-like cells could be extended. Immunocytochemical staining showed that after 10 h of neural induction, the differentiated neuron-like cells in protected group were still in a good state, and the mRNA levels of NSE and MAP-2 were increased during the induction course checked. Conclusion HSYA could improve the resistance of cells to the oxidative damage caused by BME.

Simultaneous quantitation of folic acid and 5-methyltetrahydrofolic acid in human plasma by HPLC-MS/MS and its application to a pharmacokinetic study☆ / 药物分析学报

A sensitive method based on high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed for the simultaneous determination of folic acid (FA) and its active metabolite, 5-methyltetrahydrofolic acid (5-M-THF), in human plasma. The analytes were extracted from plasma with methanol solution containing 10 mg/mL of 2-mercaptoethanol and 0.025% (v/v) ammonium hydroxide. FA and 5-M-THF were more stable after the addition of 2-mercaptoethanol and ammonium hydroxide in the sample preparation procedures of this study than they were in the previously published methods. Chromatographic separation was performed on a Hedera ODS-2 column using a gradient elution system of acetonitrile and 1 mM ammonium acetate buffer solution containing 0.6% formic acid as mobile phase. LC-MS/MS was carried out with an ESI ion-source and operated in the multiple reaction monitoring (MRM) mode. The assay was linear over the concentration ranges of 0.249-19.9 ng/mL for FA, and 5.05-50.5 ng/mL for 5-M-THF. The developed LC-MS/MS method offers increased sensitivity for quantification of FA and 5-M-THF in human plasma and was applicable to a pharmacokinetic study of FA and 5-M-THF.

Optimized medium accelerates differentiation of bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND:Studies have found that bone marrow mesenchymal stem cel s, under certain conditions, can be induced to differentiate into neurons and glial cel s, which to some extent solves the problem of the source of seed cel s. Induction methods currently used are different, and their efficiencies are not the same. OBJECTIVE:To observe the effects of different antioxidants on differentiation of rat bone marrow mesenchymal stem cel s into neuron-like cel s in vitro. METHODS:Bone marrow mesenchymal stem cel s from Wistar rats were divided into four groups:non-intervention group,β-mercaptoethanol group, retinoic acid group,β-mercaptoethanol+retinoic acid group. Changes in cel morphology and positive rate of neuron-specific enolase and microtubule-associated protein 2 were observed and detected at 5 hours, 12 hours, 1 day, 3 days, 5 days, 7 days, and 10 days after induction. RESULTS AND CONCLUSION:Except non-intervention group, bone marrow mesenchymal stem cel s in the other three groups were gradual y becoming spindle-shaped, and gave birth to many smal protrusions that were interconnected into a network, showing neuron-like cel morphology. Immunocytochemical staining showed that the efficiency of theβ-mercaptoethanol+retinoic acid group was the highest at 10 days after induction, and the positive rates of neuron-specific enolase and microtubule-associated protein 2 were 71.63%and 79.72%, respectively. The results show thatβ-mercaptoethanol can be combined with retinoic acid to accelerate the differentiation of bone marrow mesenchymal stem cel s into neuron-like cel s.

Study on the time of amounted to peak of human adipose-derived stromal cells differentiation into neural precursor cells in vitro / 中华行为医学与脑科学杂志

Objective To reseach the time point of the highest percentage of neural precursor cells derived from adipose stromal cells (ADSCs) in vitro, and to observe the ultrastructure features of neural precursor cells. Methods Used the β-mercaptoethanol to induce ADSCs to differentiate into neural precursor cells and neuron-like cells. The morphology of the uninductedcells and inducted cells were observed with inverted phase contrast microscope. The expression of nestin which was the marker of neural precursor cell in each group was detected using immunofluorescence staining method. The ultrastructural feature of cells which was induced for 3 hours were observed. Results The highest ratio of positive expression of nestin was 3 hours following induction,with the ratio ( 86.25 ± 4.82) %. There were many protuberance on the cell membrane under transmission electron microscopy.There were plenty of organelles in the neural precursor cells. The neural precursor cells had a large size nucleus,large nucleoplasmic index, much extended chromatin,and less condensed chromatin. The nucleus had double-layer nuclear envelope, more nuclear pore on the nuclear envelope. Conclusion The time point of the highest percentage of neural precursor cells derived from ADSCs is 3 hours,and the ultrastructral feature of induced neural precursor cells confirm that cells at this time point are in a state of split active period.

Millimeter-wave Exposure Promotes the Differentiation of Bone Marrow Stromal Cells into Cells with a Neural Phenotype / 华中科技大学学报（医学）（英德文版）

t of the cells treated with BME alone (P<0.05). It was concluded that MMW exposure enhanced the induc-ing effect of BME on the differentiation of BMSCs into cells with a neural phenotype.

Screening Tests for Detecting of Bacterial Metallo-?-lactamase / 中华医院感染学杂志

OBJECTIVE To establish a simpler method for screening bacterial metallo-?-lactamase(MBL) by comparing Hodge test and double-disk synergy tests(DDSTs),and using 5 inhibitors such as sodium citrate,EDTANa2,EDTAK2,sodium dimercaptopropane sulfonate and mercaptoethanol.METHODS In Hodge test,5 inhibitors of different concentration were added to the imipenem(IMP) disks,the sizes of the inhibition zones were compared.Disk containing IMP and disk containing a metallo-?-lactamase inhibitor were used in DDSTs.The optimal inhibitors concentration and edge-to-edge distance between the two disks were selected.The performances of the Hodge test and the DDSTs were also compared.RESULTS Among the metallo-?-lactamase inhibitors used in this study,1∶15 diluted mercaptoethanol gave the most reproducible and the clearest results when a filter disk containing mercaptoethanol was placed near the IMP disk with the edge distance of 3 mm.CONCLUSIONS Mercaptoethanol-IMP DDSTs are simple,convenient and sensitive methods for screening MBL.

Effect of beta-mercaptoethanol or epidermal growth factor supplementation on in vitro maturation of canine oocytes collected from dogs with different stages of the estrus cycle

Supplementation of beta-mercaptoethanol (beta-ME) in in vitro maturation (IVM) medium was shown to improve embryo development and quality in several species. Epidermal growth factor (EGF) was also shown to improve IVM of human oocyte and embryo development after in vitro fertilization (IVF). The effect of these two compounds were suggested to be mediated through the synthesis of glutathione (GSH) which is known to play an important role in protecting the cell or embryos from oxidative damage. Thus, it is suggested that supplementation of canine IVM medium with beta-ME or EGF may be of benefit due to its positive role in IVM of various mammalian oocytes and embryo development, including cattle, pigs, rodents and humans. This study investigates the effect of ovarian estrus stage on canine oocyte quality and supplementation of medium with beta-ME or EGF on IVM of canine oocytes. As results, a significantly higher percentage of oocytes progressed to metaphase II (MII) stage in 50 or 100 microM of beta-ME supplemented oocytes collected from the follicular stage. The maturation rate to metaphase I (MI) stage was also significantly higher in oocytes collected from follicular stage and cultured with 25 or 100 microM compared to other experimental groups. After IVM culture, oocytes recovered from dogs with the follicular stage and matured in TCM-199 supplemented with 20 ng/ml EGF yielded better oocyte maturation to MII phase compared to other groups. Taken together, supplementation of beta-ME (50 or 100 microM) or EGF (20 ng/ml) improved IVM of canine oocytes to MII stage.

The related gene expression in mesenchymal stem cells differentiating to neuron-like cells / 中国病理生理杂志

AIM: To analyze neuron-related gene expression before and after mesenchymal stem cells (MSCs) differentiating into neuron-like cells. METHODS: MSCs were induced to neuron-like cells with ?-mercaptoethanol. Before inducement and at 8 h after inducement, the total RNA was extracted, then the expression of microtubule-associated protein-2 (MAP-2), growth-associated protein-43 (GAP-43), NSE, nestin and neurofilament (NF) mRNA were detected with RT-PCR. RESULTS: NSE mRNA expressed before and after inducement, MAP-2, GAP-43, nestin and NF mRNA only expressed after inducement. CONCLUSION: The differentiation of MSCs into neuron-like cells may be related to MAP-2, GAP-43, nestin and NF expression. [

DETERMINATION OF INTERCELLULAR FREE Ca~(2+) CONCENTRATION IN TH E CULTURED CELLS FROM SCHISTOSOMA JAPONICUM BY USING FURA-2/A M / 中国血吸虫病防治杂志

0 05), being ( 187 0?10 7) nmol/L in average. Moreo ver, it was not significantly different from the cells cultured at day 0, ei ther . However, [Ca 2+ ]i increased significantly(P

A STUDY ON INDIRECT PEROXIDASE ANTIBODY TEST OF DETECTING IgM ANTIBODIES TO EPIDEMIC HEMORRHAGIC FEVER / 西安交通大学学报(医学版)

The immunoporoxidase staining technics (IPST) was used in 37 patients with epidemic hemorrhagic fever (EHF) for detecting the specific IgM antibodies to EHF and compared with the method of IFA-IgG. The sera obtained at the early stage of the disease were ana- lyzed. In two cases, the results were negative by using the method of IPA-IgM antibodies. In 35 cases, the results were significantly higher than those by means of the method of IFA-IgG. But, the sera obtained at the late stage showed no difference between the two methods. The inhibition test and 2-ME tolerance tcst showed that the antibodics detected by the IPST technics were specific IgM ones, which could be found in the patient's serum on the second day of onset. The results showed that the IPA-IgM method is sensitive and highspecific and the results can be read under the light microscope. Thus, this method is convenient for the country doctors to use.