Simultaneous determination of lacosamide and perampanel concentration in human plasma by LC-MS/MS / 中国药房

OBJECTIVE To establish a method for simultaneous determination of two third-generation anti-epileptic medicines such as lacosamide and perampanel in human plasma and apply this method in clinical practice. METHODS Using clozapine as internal standard， the concentrations of lacosamide and perampanel of plasma samples in 10 epileptic patients were determined by LC-MS/MS after protein precipitation with acetonitrile and dilution with acetonitrile-water （20∶80，V/V）， and the plasma minimum concentrations were obtained by dilution of multiple. The determination was performed on Welch Ultimate XB-C18 column， with mobile phase A consisted of 10 mmol/L ammonium formate and mobile phase B consisted of methanol-acetonitrile-isopropanol （0.2% formic acid） mixed solution （7∶1.5∶1.5， V/V/V） for gradient elution at the flow rate of 0.4 mL/min. The column temperature was set at 40 ℃ ， and the sample size was 5 μL. The electrospray ion source and multi-reaction monitoring mode were used for positive iron scanning. The ion pair used for quantitative analysis of lacosamide， perampanel and internal standard were m/z 251.2→ 144.1， m/z 350.2→219.2 and m/z 327.2→270.0， respectively. RESULTS The linear ranges of lacosamide and perampanel were 0.001 25-0.125 μg/mL（r＞0.99）， 0.037 5-3.75 ng/mL （r＞0.99）； the limits of quantification were 0.001 25 μg/mL and 0.037 5 ng/mL， respectively. The precision and accuracy within and between batches， extraction recovery rate， matrix effect， and stability all met relevant requirements. The minimum concentrations of lacosamide in No. 1-5 patients were 5.3-12.2 μg/mL， and the minimum concentrations of perampanel in No.6-10 patients were 208-510 ng/mL， respectively. CONCLUSIONS The established method is simple， rapid and suitable for the therapeutic drug monitoring of lacosamide and perampanel.

Frequency survey of blood group PP1PK in Chinese Lahu Population / 中国输血杂志

【Objective】 To investigate the frequency of P1PK and GLOB blood group in Chinese Lahu population and their genetic status, so as to provide data support for the safety of blood transfusion and give advice and transfusion guidance for pregnant women. 【Methods】 Unrelated individuals of Chinese Lahu population were randomly selected for serological identification of P1PK and P blood group and gene sequencing analysis. The frequency of P1PK and GLOB blood group were analyzed. 【Results】 Six cases of anti-PP 1Pk(formerly known as anti-Tja) negative blood type were identified as the rare P1-PK-P- blood type (formerly known as Tja- blood type or p blood type, hereinafter referred to as p blood group) from 300 Lahu population, with phenotypic frequency of p blood group in P1PK and GLOB blood group system in Lahu population at 2.0%(6/300). 【Conclusion】 The phenotypic frequency of blood group p in Lahu population was significantly higher than that in Europe (5.8 persons per million) and Hong Kong, China (1 person per million),indicating significant ethnic specificity and regional ethnic differences.

Preparation of human monoclonal anti-C cell line from peripheral blood B lymphocytes of D--donor / 中国输血杂志

【Objective】 To establish human hybridoma cell lines, secreting monoclonal antibody against antigens of Rh blood system, from a donor with rare D--phenotype. 【Methods】 Peripheral blood B lymphocytes of an O type female donor, lacking C/c/E/e antigens on her erythrocyte, were transformed with Epstein-Barr virus (EBVs). EBVs were harvested from the cultural supernatant of B95-8 cells. The transformed lymphoblastoid cell line (LCL) secreting antibodies to C antigens were picked up and then hybridized with the myeloma SHM-D33 using electric fusion technique. Hybridoma cells were selected by HATD-Ouabain(HOTD)(Hypoxantine, Aminopterin, Thymidine, 2-Deoxycytide, and Ouabain)culture medium, microplate antibody screening and limited dilution subcloning. The monoclonal antibody was assayed by serological test and was confirmed by flow cytometry (FCM). 【Results】 From the cultural supernatant of D--peripheral blood transformed B lymphocytes, 3A6-C6, which agglutinated with R1R1(DCe/DCe)O-type RBCs but not with R2R2(DcE/DcE)O-type RBCs, was screened and preliminarily identified as anti-C. We established a hybridoma cell line secreting anti-C immunoglobulin M from B cells of D--individual successfully after hybridization with SHM-D33 myeloma cells. 【Conclusion】 The study had laid the groundwork for future research and development of human monoclonal antibodies against Rh antigens of RBC in future for diagnosis and screening purpose.

Quantitative detection of red blood cell antibody-mediated complement activation / 中国输血杂志

【Objective】 To construct an in-vitro model of erythrocyte antibody-mediated complement activation, and establish quantitative detection methods based on flow cytometry and spectrophotometry, so as to explore the correlation of anti-body titers and complement activation speed, and provide a methodological basis for studying the adverse transfusion reactions of anti-body mediated complement hemolysis. 【Methods】 Mouse monoclonal antibody that recognized human C3b and fluorescent secondary antibody were used to label C3b fragments on erythrocytes, and the deposition of C3b fragments after complement activation was detected by flow cytometry. The absorbance at 540 nm of the supernatant in the complement activation reaction system was measured by spectrophotometry as the amount of hemoglobin released was related to the absorbance. 【Results】 The complement activation system was constructed according to the ratio of 3% red blood cell suspension (mixed for 6 people) 1∶anti-Tja 1∶complement 2. The repeatability was good (P value>0.05) as different red blood cell mixtures had been used to repeat the detection reaction system. When using 32×, 64× and 128× dilutions of anti-Tja mediated complement activation, the deposition of C3b fragments has been detected by flow cytometry at 30 s, 1 min and 2 min, respectively, and MFI peaked at 5 min, 10 min and 30 min, respectively. No obvious hemolysis has been observed within 1.5 h. 【Conclusion】 In vitro model of anti-Tja-mediated complement activation demonstrates the speed of complement activation is related to the concentration of antibody. At a certain antibody concentration, the speed of complement activation has been slowed down, and no obvious hemolysis observed.

The clinical serological characterization of cisAB and B(A) blood groups / 中国输血杂志

【Objective】 To determine the rare ABO blood subgroups rapidly and ensure the blood transfusion safety of five patients by a series of serological tests and family investigation, as their preliminary serological results of ABO blood grouping was inconsistent. 【Methods】 ABO blood grouping, antibody screening and Coombs′ tests were performed by the routine serological methods, including manual tube and automatic blood group analyzer, which had matched micro-column gel cards from Diagnostic Grifols. Polymerase chain reaction (PCR) was used to amplify the 6 and 7 exons as well as their adjacent intron region of ABO gene. The patients and their relatives′ ABO blood group and subgroup were analyzed and identified through the comparison with serological phenotype database of ABO blood group. The products of PCR were sequenced directly, and the gene mutation was identified through the comparison with the Blood Group Antigen Gene Mutation Database. 【Results】 Whether micro-column gel cards or manual tube test, the forward and reverse tests of serological grouping were not supported by each other on the five patients′ ABO blood grouping. The forward tests of patients No.1~3 showed AwB phenotype and the reverse tests showed B group. No.4 patient was the forward ABw phenotype and the reverse A group, and No.5 patient was the forward normal AB phenotype and the reverse B group, respectively. All of 5 patients′ Rh C/D/E blood grouping showed clearly; the IDT test and antibody screening result of patient No.1 was positive, while the antibody screening result of patient No.4 was negative. 【Conclusion】 The blood group serological characterization of patient No.1~3 met B(A) blood group, and patient No.4~5 met CisAB blood group. These tests can make a preliminary diagnosis of blood group phenotype, which are verified correctly via blood group genotype.

Application of MALDI-TOF MS in clinical difficult blood group typing / 中国输血杂志

【Objective】 To explore the application of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) in the genotyping of difficult blood typing samples, and to provide evidence for clinical blood transfusion. 【Methods】 Three ambiguous blood group samples, submitted to Shanghai Blood Center by Shanghai regional hospitals, were studied, of which Sample1 included the proband and his parents. Serological methods were used to perform blood group typing, direct antibody test, unexpected antibody screening and identification test. Blood group genotyping was performed by using the MALDI-TOF MS detection systeme stablished in our laboratory. Sanger sequencing was used to confirm gene mutation sites, and serological or flow methods were used to verify specific samples′ phenotype. 【Results】 Serological results indicated the existence of antibodies against high frequency antigens in sample 1 (including proband and her mother), 2 and 3. The genotyping results of MALDI-TOF MS showed that the proband of sample 1 was Di(a+ b+ ), her father was Di(a-b+ ), her mother was Di(a+ b-), sample 2 was p, and sample 3 was Jr(a-). Sequencing results of three samples were consistent with mass spectrometry typing results. Serological results showed that sample 2 had a p phenotype. The flow cytometry results suggested that sample 3 had a Jr(a-) phenotype. 【Conclusion】 For the first time, we applied MALDI-TOF MS technology to blood type genotyping of ambiguous clinical samples in China. Compared with other genotyping methods such as PCR-SSP, MALDI-TOF MS has the advantages of rapid detection, high throughput and high specificity, which would contribute to identification of difficult blood typing samples in the future, as well as rare blood group screening.

Distribution of ABO, Rh, Mn, P, h blood groups and gene frequency in Lahu ethnic minority in Yunnan / 中国输血杂志

【Objective】 To understand the distribution and gene frequency of main red blood cell blood groups in Lahu ethnic minority and analyze the genetic characteristics of Lahu people. 【Methods】 1) ABO forward and reverse typing had been performed by microplate method; 2) Rh, MN, H, P1Pk and Mur antigen were tested by the tube method. If the ABO forward and reverse typing were incompatible, the tube method was used for confirmation. 【Results】 The distribution characteristics of blood group and gene frequency in Lahu ethnic minority were as follows: B>O>A>AB for ABO, with genotype frequency as p 11.1%, q 27.5% and r 61.4%; the frequency of Rh genotype was CDe 83.3%, cDE 12.0%, cDe 2.42%, CDE 2.32%, CdE 0%, Cde 0%, cdE 0% and cde 0%; M > MN>N for MN blood group, with genotype frequency as M 75.26% and N 24.74%; P1

Baicalin protects against 17α-ethinylestradiol-induced cholestasis via Sirt1/HNF-1α/FXR pathway / 中国药理学与毒理学杂志

OBJECTIVE Baicalin is a major flavonoid component of Scutellaria baicalensis, and has been used in the treatment of liver diseases for many years. However, the role of baicalin in estrogen-induced cholestasis (EIC) remains to be elucidated. This present study explored the protective effect of baicalin against estrogen-induced liver injury and further elucidated the mechanisms involved both in vivo and in vitro. METHODS We conducted a series of experiments using 17α-ethinylestradiol (EE) induced cholestatic rats and cultured HepG2 cells. Serum, bile, and liver samples were collected for biochemical and histological analyses. Bile acid composition in liver was analyzed by LC-MS/MS. The mechanisms underlying the hepatoprotective of baicalin were investigated by RT-PCR, Western blotting analyses and immunohistochemistry. RESULTS Baicalin showed obvious hepatoprotective effects in EIC rats by reducing serum bio?markers and increasing the bile flow rate, as well as by alleviating liver histology and restoring the abnormal composition of hepatic bile acids (BAs). In addition, baicalin protected against EE induced liver injury by up-regulation of the expres?sion of hepatic efflux transporters and down-regulation of hepatic uptake transporters. Furthermore, baicalin increased the expression of hepatic BA synthase (CYP27A1) and metabolic enzymes (Bal, Baat and Sult2a1) in EIC rats. We showed that baicalin significantly inhibited hepatic inflammatory responses in EIC rats through reducing elevated levels of TNF-α, IL-1β, IL-6 and NF-κB. Finally, we confirmed that baicalin maintains BA homeostasis and alleviates inflamma?tion through Sirt1/HNF-1α/FXR signaling pathway. CONCLUSION Baicalin protects against estrogen-induced cholestatic liver injury, and the underlying mechanism involved is related to activation of the Sirt1/HNF-1α/FXR signaling pathway.

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.

Genetic polymorphism and antibody screening of RhD negative Chinese Han population in Jiayuguan / 中国输血杂志

【Objective】 To investigate and analyze the polymorphism of RHD gene in RhD-negative population in Jiayuguan using molecular biological technique, so as to accurately identify RhD-negative individuals, and formulate individualized transfusion strategies. 【Methods】 The RhD negative voluntary blood donors and patients (mainly pregnant women) were recruited. After informed consent, history of blood transfusion and pregnancy of them were investigated, and samples were collected for negative D confirmation, gene sequencing as well as antibody screening and identification. 【Results】 Among the 96 samples, 73 cases were RHD gene deletion, 18 RHD*01EL.01(17 RHD1227A homozygous type and 1 RHD1227A heterozygous type), 2 weak RHD*15 type (845G/A), 1 partial D type, i. e. RHD-CE(7) -D heterozygous allele, and 2 RHD*01N.16 variant. Antibody was detected out in 4 cases, among which 2 were positive for anti-D, 1 anti-D plus anti-E, and 1 anti-Dia. 【Conclusion】 The proportion of DEL gene in RhD negative Chinese Han population in Jiayuguan is slightly lower than that in general Chinese Han population. No anti-D or RHD-HDN was observed in DEL type women due to multiple pregnancy or delivery of D positive newborns.

-D- phenotype screening by polybrene method / 中国输血杂志

【Objective】 To develop a novel screening reagent for -D- phenotype preliminary screening based on the difference in RhD antigen expression level of -D- phenotype and normal RhD phenotype. 【Methods】 RhD antigen expression of -D-phenotype and Rh D-- gene carrier were detected by flow cytometry. By adjusting the concentration of polybrene in the screening system, the red blood cells with high RhD antigen expression level agglutinated, and the preliminary screening of the -D-phenotype and its gene carriers was realized. 【Results】 According to the quantitative results of immunofluorescence intensity (MFI) analysis by flow cytometry, the expression level of RhD antigen in -D- phenotype cells (284 360±16 698, n=3) was about 3 times normal RhD positive cells (98 642±35 908, n=9)(P<0.01), while RhD antigen expression level of RhD-- gene carrier (181 109±39 455, n=4) was about 2 times normal RhD positive cells(P<0.01). RhD antigen expression (144 538±227 445, n=7) of the positive cells screened by 15 μL 3% fresh red blood cell suspension and screening system 35 μL (1 μL IgG anti-D, 29 μL polybrene polybrene, and 5 μL low ionic strength solution) was about 1.5 times normal RhD positive cells. 【Conclusion】 The polybrene preliminary screening system, which can be used for high-throughput screening of -D- phenotype, is a reliable technical method for frequency study of this phenotype.

Changes in the proliferation and angiogenesis of human dental pulp stem cells after treated with prostaglandin E1 combined with basic fibroblast growth factor / 中国组织工程研究

BACKGROUND: Prostaglandin E1 and basic fibroblast growth factor can promote the proliferation of human dental pulp stem cells, but the effects of their combinations on the proliferation of human dental pulp stem cells and angiogenesis are unknown. OBJECTIVE: To investigate the effects of prostaglandin E1 combined with basic fibroblast growth factor on the proliferation and angiogenesis of human dental pulp stem cells. METHODS: (1) Human dental pulp stem cells were isolated and cultured in vitro. After detection and identification of surface markers, prostaglandin E1 and basic fibroblast growth factor at concentrations of 5, 10, 20, 50 and 100 μg/L were used to treat human dental pulp stem cells in vitro. The untreated cells served as control group. The cell proliferation was detected by cell counting kit-8 assay, and the optimum drug concentration and time of drug action were screened. (2) The in vitro cultured human dental pulp stem cells were divided into four groups: blank control group, prostaglandin E1 group, basic fibroblast growth factor group and combination group. The cell proliferation was detected by cell counting kit-8 assay. Human dental pulp stem cell conditioned medium was extracted. The levels of vascular endothelial growth factor and endostatin in the culture medium were detected by ELISA. The in vitro tubular formation ability of human umbilical vein endothelial cells after culture in conditioned medium was tested by tubule formation experiment. RESULTS AND CONCLUSION: The optimum concentration of prostaglandin E1 and basic fibroblast growth factor was 20 μg/L, and the optimum time of action was 2 days. Compared with the blank control group, the relative proliferation rate, level of vascular endothelial growth factor and the angiogenesis ability of human umbilical vein endothelial cell in vitro in the prostaglandin E1, basic fibroblast growth factor and combination groups were significantly increased (P < 0.05), while the level of endostatin was significantly decreased (P < 0.05). All above index levels in the combination group were significantly superior to those in the prostaglandin E1 and basic fibroblast growth factor groups (P < 0.05). In summary, prostaglandin E1 combined with basic fibroblast growth factor can promote the proliferation of human dental pulp stem cells and enhance the tubular formation ability of human umbilical vein endothelial cells in vitro.

Effect and Mechanism of Pilose Antler Different Components on Bone Tissue of Ovariectomized Osteoporosis Model Rats / 中国实验方剂学杂志

Objective:To explore the effect and mechanism of pilose antler different components on the bone tissue of ovariectomized osteoporosis model rats and ascertain the material basis of pilose antler. Method:fifty-six SD rats were divided randomly into seven groups:normal group,model group,Xianling Gubao group(468 mg·kg-1),Bujiale group(80 mg·kg-1),polysaccharide group(50 mg·kg-1),polypeptides group(175 mg·kg-1),polysaccharide and polypeptide mixture group(50 mg·kg-1+175 mg·kg-1). Osteoporosis mode was established through ovary resection of female rats,meanwhile,the rats were given different components of pilose antler for consecutively 12 weeks. Subsequently, using absorptiometry to measure the rats' bone mass density. The activities of bone alkaline phosphatase(BALP),osteocalcin (OT),bone morphogenetic protein2(BMP-2),Smad1,Smad5,Runt-related transcription factor 2 (RUNX2) were detected by enzyme-linked immuno sorbent assay (ELISA). The expression of BMP-2,Smad1,Smad5,Runx2 protein was examined by Western blot and Real-time polymerase chain reaction (Real-time PCR). Morphological assay for bone tissue were detected by htoxylin eosin(HE) staining. Result:After 12 weeks, Compared with the normal group, the osteoporosis model group showed significantly decrease in bone mineral density(PPPConclusion:Pilose antler different components has therapeutic effect on ovariectomized osteoporosis model rats.The mechanism may be related to up-regulat the expression of BMP-2/Smad1,Smad5/Runx2 signal pathways.

Mechanism of Calculus Bovis Sativus in inhibiting hepatocyte lipid deposition based on serum pharmacology / 中国中药杂志

The aim of this paper was to investigate the molecular mechanism of Calculus Bovis Sativus( CBS) in alleviating lipid accumulation in vitro by serum pharmacology. The CBS-containing serum of mice was obtained by serum pharmacology method to evaluate its effect on the proliferation of LO2 hepatocytes. The lipid reducing effects of CBS-containing serum through Nrf2 was evaluated by fructose-induced LO2 hepatocyte steatosis model,nuclear factor erythroid 2 related factor 2( Nrf2) agonist oltipraz combined intervention,cell oil red O staining and intracellular triglyceride( TG) content. The effects of CBS-containing serum on lipid peroxidation and hepatocytes apoptosis were evaluated by reactive oxygen species( ROS) and apoptosis assay,respectively. Real-time quantitative polymerase chain reaction( PCR) was used to detect the relative expression of lipid synthesis-related genes and apoptosis-related genes.RESULTS:: showed that CBS drug-containing serum had no significant effect on LO2 hepatocyte proliferation. As compared with the model group,CBS-containing serum could effectively reduce the formation of lipid droplets in fructose-induced LO2 hepatocytes,significantly reduce intracellular TG and ROS levels,and significantly reduce hepatocyte apoptosis rate( P < 0. 05). As compared with the model group,carbohydrate responsive element binding protein( ChREBP),sterol regulatory element binding protein-1 c( SREBP-1 c),fatty acid synthase( FAS),acetyl-CoA carboxylase 1( ACC1),stearoyl-CoA desaturase 1( SCD1),Bax and caspase-3 mRNA levels were significantly reduced in CBS drug-containing serum treatment group( P<0. 05). All of the above effects could be reversed by oltipraz.In conclusion,CBS-containing serum can significantly inhibit the fructose-induced LO2 liver fat deposition,and the mechanism may be related to reducing intracellular ROS level through the Nrf2 pathway and improving intracellular peroxidation state to reduce apoptosis.

A evaluation for uncertainty analysis of the verification result of ELISA analyzer(microplate reader) / 中国医学装备

Objective: To analyze various factors that affect the verification result in the verification by using ELISA analyzer and assess the accuracy of verification results of ELISA analyzer. Methods: Through the comparison of verification results and the calculation for the repeatability of measurement standard, the accuracy of standard device was analyzed and verified. Results: Through analyzed the effect of repeatability of operation and standard device, the factors that affected verification results were eliminated. And the accuracy of verification results were guaranteed. Conclusion: In the process of verification for ELISA analyzer, operators should pay attention to the setting of the verification instrument. And the operation regulation can ensure the accuracy and reliability of the measured value of spectrophotometer. Besides, operators also should pay attention to the analysis of verification results and confirm whether the repeatability of the verification results can meet the requirement of practice.

Relationship Between Distribution of Oral Ulcer and Zang-fu Viscera Syndrome Differentiation / 广州中医药大学学报

Objective To study the relationship between the distribution of oral ulcer and the syndrome differentiation of Zang-fu viscera. Methods We collected the data of 65 oral ulcer patients admitted in Stomatology Department,the First Affiliated Hospital of Guangzhou University of Chinese Medicine from January of 2015 to July of 2016. The data of emotion,diet,sleep,history of digestive tract disease,and family history of the patients with tongue ulcer and the patients with labio-buccal ulcer were compared. The relationship of the distribution of labio-buccal ulcer,tongue-tip ulcer,and ulcer of lateral part of the tongue with Zang-fu viscera syndrome differentiation was analyzed by statistical software SAS 9.2. Results(1)Tongue ulcer group had higher incidence of anxiety and higher percentage of history of digestive tract disease than labio-buccal ulcer group(P<0.01,P<0.05). (2)Labio-buccal ulcer mostly had a correlation with spleen deficiency blended with dampness or gastrointestinal accumulating heat, the tongue-tip ulcer mostly had a correlation with heart-fire flaring up, and the ulcer of lateral part of tongue mostly had a correlation with pathogenic fire derived from liver-qi stagnation, the difference among the 3 groups being statistically significant(P < 0.01). Conclusion The distribution of oral ulcer has a correlation with Zang-fu viscera syndrome differentiation.

The application of dynamic high-frequency ultrasound in the diagnosis of Zenker diverticulum / 中国医学装备

Objective:To explore the diagnostic value of high-frequency ultrasound for Zenker diverticulum.Methods: 15 patients who were suspected as Zenker diverticulum through the diagnosis of using high frequency ultrasound were analyzed in the research. Their appearances of ultrasound were summarized, and these results were compared with barium meal at upper gastrointestinal tract and results of postoperative pathology, respectively.Results: In 15 patients, 4 cases were confirmed by adopting barium meal at upper gastrointestinal tract, and 11 cases were confirmed by adopting postoperative pathology. All of lesions in 15 cases were located on the back of left side of the thyroid gland, and there were 3 kinds of sonographic appearance. The first kind was equal echo lesion, and there were spots and schistoses without echo inside lesion, their form showed a semicircle shape. The second kind was hypoechoic, the form showed irregularity or semi cyclic annular, and the border was clear, and there were strong echogenic spots. The third kind was hyperechoic lesions, and the strong echo were movable with morphological changing after the slight pressure of search unit, and slim half ring low echo wall can be seen indistinctly around lesions.Conclusion:High-frequency ultrasonography is a convenient, rapid and non-invasive method for the diagnosis of Zenker diverticulum, and it is helpful to grasp its ultrasonogram characteristic and examination method in early detection of disease, avoiding misdiagnosis and missed diagnosis. Therefore, it has important clinical value.

Study on the Specificity Enrichment of Hepatoma Cell Monoclonal Antibody Against Hepatocellular Carcinoma and Gastric Cancer / 现代检验医学杂志

Objective To investigate whether the combination of 4-E9 immunomagnetic beads and epithelial adhesion mole-cules(Epacm)beads can enhance the enrichment efficiency of MCF-7,BEL-7402 and BGC-823 cells.Methods A mono-clonal antibody was prepared and ligated with magnetic bead by a biotin and a streptavidin to prepare an immunomagnetic beads.The enrichment rate of MCF-7,BEL-7402 and BGC-823 cells was detected by the combination of two kinds of immu-nomagnetic beads and two kinds of immunomagnetic beads.Results The encapsulation rate of 4-E9 immunomagnetic beads was 57.8%,and the encapsulation rate of Epcam immunomagnetic beads was 65.8%.4-E9 immunomagnetic beads on MCF-7,BGC-823 and BEL-7402 cell capture rate was(44±5.33)%,(71±11.33)% and(78.3±8.46)% respectively.Epcam immunomagnetic beads on BGC-823,BEL-7402 and MCF-7 cell capture rate was(55.5±8.67)%,(78.88±13.11)% and (84.31±6.83)% respectively.The combination of two kinds of immunomagnetic beads on BGC-823,BEL-7402 and MCF-7cell capture rate was(80.4±8.33)%,(85.125±6.77)% and(93.23±4.33)% respectively.Joint beads group compared with 4-E9 immunomagnetic beads on BGC-823,BEL-7402 and MCF-7 cell enrichment rate were statistically significant(P=0.03,0.03,0.04),and joint beads group compared with 4-E9 immunomagnetic beads on BGC-823,BEL-7402 and MCF-7 cell enrichment rate were statistically signific(P=0.04,0.03,0.04).Conclusion The combination of two kinds of immunomag-netic beads can significantly improve the enrichment efficiency of Epcam immunomagnetic beads on BEL-7402,BGC-823 and MCF-7 cells.4-E9 antibody enrichment of circulating tumor cells may have some clinical value.

Improvement of medical equipment quality control management module / 医疗卫生装备

Objective To improve medical equipment quality control management module to report medical equipment adverse events,evaluating medical equipment,managing medical metrology certificate and etc.Methods The module with four functions was developed with the original equipment management information system as well as the list structures of eq_measuring and eq_measuring_detailed and Microsoft Visual Studio 2010.Rwesults The system realized the management of adverse events,evaluation,scrapping and certificate,and could be used to master the quality of in-use equipment based on data analysis.Conclusion The system behaves well in stability and usage,and thus is worthy promoting for enhancing medical equipment quality coutrol and management.

Detection of Thrombin Using Double Quenching Molecular Beacon / 分析化学

A double quenching molecular beacon ( MB) with simple structure was designed based on organic quencher and G bases, and a simple detection method for thrombin was developed using this MB. In this MB, FAM and BHQ-1 were selected as fluorophore and organic quencher, three continuous nucleotides with G base were connected with BHQ-1, and the loop of MB was designed as a nucleic acid aptamer of thrombin. In the absence of thrombin, the MB was in the stem-loop structure, the fluorophore FAM was close to BHQ-1 and G bases, the fluorescence of FAM was dual quenched by BHQ-1 and G bases, and the fluorescence signal of FAM was very weak. In the presence of thrombin, MB specifically bound thrombin and formed a G-quadruplex structure. The stem-loop structure of the MB was destroyed, and FAM was separated with BHQ-1 and G bases, leading to recovery of fluorescence of FAM. Under the optimal conditions, the fluorescence intensity of FAM exhibited a good linear relationship with concentration of thrombin in the range of 0. 4-40. 0 nmol/L, and regression equation was △I=24. 63C (nmol/L)+ 13. 06 (R2 =0. 9972) with the detection limit of 0. 18 nmol/L (3σ, n=9). The average recoveries of this method in serum samples were 96. 3%-98. 7%, which indicated that the method had high accuracy.