Value of drug resistance and homology of Acinetobacter baumannii in tracing the source of nosocomial infection in Jianyang, Sichuan / 中国热带医学

@#Abstract: Objective　To analyze the drug sensitivity and the carrying of carbapenem resistant gene of Acinetobacter baumannii isolated from clinical patients and clinical objects, and analyze the homology of strains to provide support for the control of nosocomial infection. Methods　A total of 38 strains of Acinetobacter baumannii isolated from patients and clinical objects surface were collected from January 2019 to August 2020. The antimicrobial susceptibility was tested by the minimum inhibitory concentration method. In addition, the resistance related genes were detected by polymerase chain reaction method, and homology analysis was performed by enterobacterial repetitive Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR). Results All 34 strains of Acinetobacter baumannii isolated from Clinical patients and 4 strains isolated from clinical objects carried blaOXA-51 and imp resistance genes, neither of them carried blaVIM gene. 32 Acinetobacter baumannii carrying blaOXA-23 gene, 28 strains carrying blaTEM gene, 7 strains carrying blaOXA-58 gene. After cluster analysis, 38 Acinetobacter baumannii isolates were classified into 7 genotypes (expressed A, B, C, D, E, F, G), and cluster E and cluster G were the main clusters, containing 12 strains (12/38, 31.6%) and 18 strains (18/38, 47.4%), respectively, as the main prevalent clonal strains. Conclusions　Acinetobacter baumannii isolated from different sources have the significant differences in drug resistance and carry different resistance genes. There is no direct correlation between patients and environmental isolates of Acinetobacter baumannii belonging to different clonal strains. Also, there aren’t significant correlation between clinical patients infected with Acinetobacter baumannii.

Effect of Stains on the Purity Determination of Human Serum Albumin by Agarose Gel Electrophoresis / 中国药学杂志

OBJECTIVE: To study the effect of stains on the purity detection results of human serum albumin by agarose gel electrophoresis, thus to select appropriate stain so that the detection result of agarose gel electrophoresis method can be basically consistent with the method specified in the 2015 edition of Chinese Pharmacopoeia, cellulose acetate membrane electrophoresis method. METHODS: Acid blue, amino black and ponceau were used in agarose gel electrophoresis to detect the purity of 30 batches of human serum albumin samples, and the results were compared with those obtained by cellulose acetate membrane electrophoresis. RESULTS: Impurity bands could not be detected by ponceau stain; the results of amino black staining was significantly lower than that of cellulose acetate membrane electrophoresis. There was no statistical difference between the results of amino black staining and cellulose acetate membrane electrophoresis. The results of 25 batches of samples detected by cellulose acetate membrane electrophoresis were within the 95% confidence interval of the results detected by amino black staining. CONCLUSION: The category of stain has great influence on the detected purity of human serum albumin when using agarose gel electrophoresis. The results of amino black staining is consistent with those of the method recorded in the Chinese Pharmacopoeia in use.

Influence of cell free fetal fraction DNA for noninvasive prenatal test / 国际检验医学杂志

Objective To explore the influence of lower concentration of cell free fetal fraction DNA in maternal plasma on non-invasive prenatal test(NIPT) .Methods A total of 3240 pregnant women accepted NIPT in Foshan Maternal and Children′s Hos-pital from April ,2015 to March ,2016 were analyzed retrospectively ,and 150 samples of which were male fetus judged by Z score of Y chromosome and the cell free fetal fraction DNA were lower than 8% were selected .The cell free fetal fraction DNA were in-creased by agarose gel electrophoresis ,then conducted NIPT ,compared with the results of aneuploidy screening .Results The cell free fetal fraction DNA were increased from 5% to 9 .2% by agarose gel electrophoresis .The result of NIPT after increasing fetal fraction was consistent with it before .Conclusion Concentration of cell free fetal fraction DNA has no influence on the result of NIPT when cell free fetal fraction DNA is above 5% .

Influence of cell free fetal fraction DNA for noninvasive prenatal test / 国际检验医学杂志

Objective To explore the influence of lower concentration of cell free fetal fraction DNA in maternal plasma on non-invasive prenatal test(NIPT) .Methods A total of 3240 pregnant women accepted NIPT in Foshan Maternal and Children′s Hos-pital from April ,2015 to March ,2016 were analyzed retrospectively ,and 150 samples of which were male fetus judged by Z score of Y chromosome and the cell free fetal fraction DNA were lower than 8% were selected .The cell free fetal fraction DNA were in-creased by agarose gel electrophoresis ,then conducted NIPT ,compared with the results of aneuploidy screening .Results The cell free fetal fraction DNA were increased from 5% to 9 .2% by agarose gel electrophoresis .The result of NIPT after increasing fetal fraction was consistent with it before .Conclusion Concentration of cell free fetal fraction DNA has no influence on the result of NIPT when cell free fetal fraction DNA is above 5% .

Analysis and significance of results of hemoglobin electrophoresis in 7 652 cases / 国际检验医学杂志

Objective To investigate the detection rates and types of hemoglobinopathy in Xiaolan district of Zhongshan city to provide the basis for prepotency .Methods 7 652 pregnant women with pregnancy detection in our hospital from June 2010 to De‐cember 2013 were selected and performed the electrophoresis by the Hydrasys automatic hemoglobin electrophoresis system .The hemoglobin components were scanned out and performed the statistical analysis .Results Among 7 652 samples ,1 057 cases of sus‐pected hemoglobinopathy were screened out with the detection rate of 10 .36% .Among them ,511 cases(6 .68% ) were Hb A2 3 .5% (suspected β‐thalassemia) ,the detection rate of Hb A2 >3 .5% was lower than that of Hb A2 < 2 .0% ,the difference had statistical significance(P< 0 .05) .11 cases (0 .14% ) were Hb C or E ,9 cases (0 .12% ) were Hb J or K ,8 cases (0 .10% ) were Hb H ,7 cases (0 .09% ) were Hb Bart′s ,1 case was Hb CS ,2 cases were Hb G and 1 case was Hb D ,which accounted for 0 .04% .Conclusion Hemoglobin electrophoresis can screen hemoglobinopa‐thy effectively .The hemoglobinopathy screening in this area should be conducted as soon as possible ,which has an important signif‐icance to prepotency .

Synthesis of nonviral gene carrier poly (L-glutamic acid)-graft-polypropylenimine and its complexation stability with DNA / 中国药学杂志

OBJECTIVE: To synthesize nonviral gene carrier poly(L-glutamic acid)-graft-polypropylenimine(MP-g-PPI) and investigate the capability of the polymers to condense DNA and the stability of MP-g-PPI DNA complexes.

Polyclonal all immune globulin hematic disease laboratory appraisal analysis discussed in this paper / 国际检验医学杂志

Objective To explore the clinical diagnostic value of polyclonal all immunoglobulin.Methods The specimens of pa-tients were simultaneously tested and identified by quantitative immunoglobulins,Immunofixation electrophoresis of serum and u-rine,urine protein electrophoresis,and other ways.Results From 1 patients with rheumatoid arthritis were detected the serum IgG, M,A,KAP and LAM,and urine KAP and LAM,at the same time show the increment of the polyclonal polyclonal all immune glob-ulin hematic disease.Conclusion Polyclonal all immune globulin hematic disease often appear in the complications of chronic in-flammation,which should be paid attention during its in clinical doctors.

Curing effects evaluation of rhizoma atractylodis macrocephalae on yeast prion[PSI+] / 中国人兽共患病学报

Investigation of the effects of rhizome atractylodis macrocephalae on yeast prion [PSI+ ]was conducted in this study .Liquid culture containing rhizome atractylodis macrocephalae aqueous extracts was applied to evaluate its therapeutic effects preliminarily .Then yeast replica plating together with Semi Denaturing Detergent Agarose Gel Electrophoresis com-bined with western blot were used to further confirm the results .It's showed that the cure rate of aqueous extracts of rhizome atractylodis macrocephalae on yease prion [PSI+ ]could reach 6% .

Application of improved gel electrophoresis to the detection of genotype polymorphisms / 中华实用儿科临床杂志

Objective To explore the traits of improved polyacrylamide gel electrophoresis (PAGE)-ethidium bromide(EB) staining in the detection of genotype polymorphisms.Methods The methods of PAGE-silver staining,agarose gel electrophoresis (AGE)-EB staining and improved PAGE-EB staining in polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to detect the μ-opioid receptor (OPRM1) A118G genotypes in case group (n =167) infants with intracranial hemorrhage (ICH),and control group(n =163) infants without ICH,to conduct a case study analysis.And the application traits of three methods were compared.Results Genotypes of OPRM1 A118G were GG (169 bp),AG (193 bp,169 bp),AA (193 bp).Both the electrophoresis methods of PAGE-silver staining and PAGE-EB could be used to detect the genotypes of OPRM1 A118G clearly in this study.There was no statistically significant difference between the resolutions of DNA fragments (P =0.884).The first method,which had 13 experiment steps,consuming 4-5 hours,involving 12 kinds of chemical reagents,and the pictures were taken with the camera,was complex,with difficult operation,more time consuming; Compared with the first method,the secondmethod was simple,which had 6 test procedures,consuming 2 hours with 8 reagents,and the pictures were taken by using an automatic gel imager.AGE-EB could not be used to detect genotypes of OPRM1 A118G.Conclusions The method of improved PAGE-EB has the advantages of fast,easy operation,and high resolution,which is worthy of wider application.

Degradation of bacterial DNA by a natural antimicrobial agent with the help of biomimetic membrane system.

The antimicrobial efficacy of methylglyoxal (MG) against several gram-negative bacteria including Escherichia coli has been reported. To determine the mechanism of action of MG, molecular interactions between lipid and MG within the liposomal membrane were also investigated. Multilamellar and unilamellar vesicles were prepared from 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). The effect of MG on DPPC liposomal membrane was studied by fluorescence spectroscopy and differential scanning calorimetry. The results indicate that MG interacts mainly with the DPPC head group that produces a significant increase in the fluidity of liposomal vesicles, which could be the cause of a fusion/aggregation effect in microbial cells. The agarose gel electrophoresis study with the genomic DNA extracted from E. coli ATCC 25922 revealed that addition of MG could completely degrade this DNA within 1 h, pointing out to their distinctly high degree of sensitivity towards MG. Further, the drug was able to cross the cell membranes, penetrating into the interior of the cell and interacting with DNA for demonstrating antibacterial activity of MG.

Técnicas electroforéticas en el HYDRASYS 2. Utilidad diagnóstica en diferentes enfermedades / Electrophoretic techniques in HYDRASYS 2. Diagnostic usability in different diseases

Se presentan los resultados de la estandarización de las técnicas de electroforesis de hemoglobina (Hb), isoenzimas de la deshidrogenasa láctica (LDH) y proteinuria en el equipo Hydrasys 2, así como el estudio de pacientes atendidos en el Instituto de Hematología e Inmunología y en otros centros hospitalarios del país. Se realizó el diagnóstico de 149 portadores de hemoglobinopatías (AS, AC, b talasemia heterocigótica, variante rápida), 60 enfermos (SS, SC, CC), 24 pacientes con a talasemia o deficiencia de hierro y se cuantificó la hemoglobina fetal a 93 casos con hemoglobinopatía S. Se determinaron los valores normales de actividad e isoenzimas de LDH en la población mediante el estudio de 50 donantes de sangre. En los pacientes con anemia drepanocítica se encontró un aumento significativo de la isoenzima 1 (p= 0,000) y disminución de isoenzimas 3 (p= 0,002). Se realizó el estudio de proteínas en orina a 8 pacientes con enfermedades hematológicas que presentaron microalbuminuria al menos en 2 ocasiones, con concentraciones ³ 0,04 g/L. En 2 pacientes el resultado fue normal; en 2 se encontraron proteínas de origen tubular; y en otros 2, proteínas de origen glomerular

We present the results of the standardization of the techniques of electrophoresis of hemoglobin (Hb), lactate dehydrogenase isoenzymes (LDH) and proteinuria in HYDRASYS 2 equipment, and the study of patients treated at the Institute of Hematology and Immunology and other hospitals in the country. 149 hemoglobinopathies carriers were diagnosed (AS, AC, b thalassemia heterozygous fast variant), 60 patients (SS, SC, CC), 24 patients with athalassemia or iron deficiency. Fetal hemoglobin was quantified in 93 cases with hemoglobinopaty S. Normal values of activity and LDH isoenzymes were determined in the population through the study of 50 blood donors. In patients with sickle cell anemia we found a significant increase in isoenzyme 1 (p=0.000) and isozyme 3 decreased (p=0.002). We performed the study of proteins in urine in 8 patients with hematologic malignancies who had microalbuminuria at least 2 times, with concentrations ³ 0.04 g / L. In 2 patients the results were normal, in 2 proteins were tubular origin, and in 2, proteins of glomerular origin

Synergistic activity of ofloxacin and ornidazole on biomedical fabrics against nosocomial pathogens.

Nosocomial infections in the hospitals disseminated from the cotton fabrics of health care professionals and patients leads to severe complications like respiratory, gastrointestinal and urinary tract infections. Since the hospital based textile materials like nylon and polyester has good surface properties, it can harbour large number of microorganisms. Hence in this study, two different antibacterial drugs showing synergistic properties were attached to different fabricsusing tocopherol acetate as a cross-linker with the aim that, treated fabric could act as barriers against transmission of challenge organisms. Inorder to decrease the drug resistant property of the nosocomial pathogens, a fluoroquinolone and a nitroimidazole compounds were mixed at suitable composition based on their synergistic behaviour. Both the compounds were modified to act as reactive dyes and were covalently bonded to the surface of nylon and polyester in order to impart antibacterial properties. The assay used for measuring antibacterial properties was based on the AATCC Test Method-100. The treated fabric was also subjected to multiple washings to determine its durability based on the AATCC Test Method-124. To determine the mode of action of these drugs, DNA of the drug exposed and unexposed challenge organisms were extracted and analysed by agarose gel electrophoresis. The difference in the number of viable bacteria after ‘0’ contact time and 18 hours contact time with treated fabrics were statistically calculated with P<0.05 considered significant.

Appraisal of Antihyperlipidemic Activities of Lentinus lepideus in Hypercholesterolemic Rats

The wild edible mushroom, Lentinus lepideus has recently been cultivated for commercial use in Korea. While the mushroom has been widely used for nutritional and medicinal purposes, the possible anti-hyperlipidemic action is unclear. The effects of dietary L. lepideus on plasma and feces biochemical and on the liver histological status were investigated in hypercholesterolemic rats. Six-wk-old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. Biochemical and histological examinations were performed. A diet containing 5% L. lepideus fruiting bodies reduced plasma total cholesterol, triglyceride, low-density lipoprotein, total lipid, phospholipids, and the ratio of low-density to high-density lipoprotein. Body weight was reduced. The diet did not adversely affect plasma biochemical and enzyme profiles. L. lepideus reduced significantly plasma beta- and pre-beta-lipoprotein, while alpha-lipoprotein content was increased. A histological study of hepatic cells by conventional hematoxylin-eosin and oil red O staining revealed normal findings for mushroom-fed hypercholesterolemic rats. The present study suggests that a diet supplemented with L. lepideus can provide health benefits by acting on the atherogenic lipid profile in hypercholesterolemic rats.

Antihyperlipidemic Effect of Dietary Lentinus edodes on Plasma, Feces and Hepatic Tissues in Hypercholesterolemic Rats

We investigated diet supplementation with shiitake mushroom fruiting bodies on biochemical and histological changes in hypercholesterolemic rats. Six-wk old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. A diet containing 5% Lentinus edodes fruiting bodies given to hypercholesterolemic rats reduced plasma total cholesterol, triglyceride, low-density lipoprotein (LDL), total lipid, phospholipids, and the LDL/high-density lipoprotein ratio by 34.33, 53.21, 75.00, 34.66, 25.73, and 71.43%, respectively. Feeding mushroom also significantly reduced body weight in hypercholesterolemic rats. However, it had no detrimental effects on plasma albumin, total bilirubin, direct bilirubin, creatinine, blood urea nitrogen, uric acid, glucose, total protein, calcium, sodium, potassium, chloride, inorganic phosphate, magnesium, or enzyme profiles. Feeding mushroom increased total lipid and cholesterol excretion in feces. The plasma lipoprotein fraction, separated by agarose gel electrophoresis, indicated that L. edodes significantly reduced plasma beta and pre-beta-lipoprotein but increased alpha-lipoprotein. A histological study of hepatic cells by conventional hematoxylin-eosin and oil red-O staining showed normal findings for mushroom-fed hypercholesterolemic rats. These results suggest that shiitake mushrooms could be recommended as a natural cholesterol lowering substance in the diet.

Hypolipidemic Activities of Dietary Pleurotus ostreatus in Hypercholesterolemic Rats

This work was conducted to investigate dietary supplementation of oyster mushroom fruiting bodies on biochemical and histological changes in hyper and normocholesterolemic rats. Six-week old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. Feeding a diet containing a 5% powder of Pleurotus ostreatus fruiting bodies to hypercholesterolemic rats reduced plasma total cholesterol, triglyceride, low-density lipoprotein (LDL), total lipid, phospholipids, and LDL/high-density lipoprotein ratio by 30.18, 52.75, 59.62, 34.15, 23.89, and 50%, respectively. Feeding oyster mushrooms also significantly reduced body weight in hypercholesterolemic rats. However, it had no adverse effects on plasma albumin, total bilirubin, direct bilirubin, creatinin, blood urea nitrogen, uric acid, glucose, total protein, calcium, sodium, potassium, chloride, inorganic phosphate, magnesium, or enzyme profiles. Feeding mushroom increased total lipid and cholesterol excretion in feces. The plasma lipoprotein fraction, separated by agarose gel electrophoresis, indicated that P. ostreatus significantly reduced plasma beta and pre-beta-lipoprotein but increased alpha-lipoprotein. A histological study of hepatic cells by conventional hematoxylin-eosin and oil red O staining revealed normal findings for mushroom-fed hypercholesterolemic rats. These results suggest that a 5% P. ostreatus diet supplement provided health benefits by acting on the atherogenic lipid profile in hypercholesterolemic rats.

DNA-UMSAgen, extracción de DNA genómica para diagnóstico molecular: método rápido y económico / UMSAgen DNA, genomic DNA extraction for molecular diagnosis: rapid and economical method

Introducción. El estudio del ácido desoxirribunocleico (DNA) es imprescindible para la medicina moderna; por ello, se ha diseñado diferentes técnicas para su extracción, cuyos costos son altos y de tiempo prolongado. En este trabajo se describe un método modificado de extracción de DNA (DNA-UMSAgen) basado en la técnica de Miller. Métodos. Las células mononucleares fueron obtenidas de sangre venosa periférica de un sujeto voluntario, para realizar 40 extracciones de DNA; 20 con el método modificado DNA_UMSAgen y otros 20 con el método clásico. Posteriormente se evaluó la concentración, calidad y utilidad de estos DNA extraidos. Resultados. La pureza del DNA extraido por el método DNA-UMSAgen es de 1,88 similar al de la técnica clásica 1,91, las concentraciones obtenidas son 20,4 ug/106 cel y 56ug/106 cel respectivamente. La evaluación por electroforesis en agarosa y la amplificación del exon 12 del gen JAK2 por PCR fue satisfactoria. Conclusión. El método DNA-UMSAgen es una alternativa de extracción de DNA genómico, rápido y económico, adecuado para paises en vias de desarrollo.

INTRODUCTIONDNA is the genetic material of the cell. Actually for its study, laboratory techniques are available that require its extraction free of impurities. This paper describes the DNA-UMSAgen method as an alternative for DNA extraction which is based on the Miller technique.METHODSThe mononuclear cells were obtained of venous peripheral blood of a voluntary subject, for to realize 40 DNA's extractions; 20 with the modified method DNA-UMSAgen and other 20 with the classic method. Later there was evaluated the concentration, quality and utility of these extracted DNA.RESULTSThe DNA purity extracted by the DNA-UMSAgen method is 1,88 similar to the classic technique 1,91, the concentration obtained were 20,4 ug/106 cells and 56ug/106 cells respectively. The evaluation by agarose gel electrophoresis and the amplification of the exon 12 of the JAK2 gen by PCR was successful.CONCLUSIONThe DNA-UMSAgen extraction method is a very acceptable fast, easy and inexpensive alternative method for underdeveloped countries for DNA extraction.

Comparison between Real-Time PCR and Agarose Gel Electrophoresis for DNA Quantification / 대한진단검사의학회지

BACKGROUND: Real-time polymerase chain reaction (PCR) is generally regarded as a very accurate and time-saving method, but it is expensive to run. We evaluated the reliability of an inexpensive and a researcher-friendly gel electrophoresis-based PCR method for the quantification of mRNA, and the results were compared with those obtained by real-time PCR. METHODS: We compared the results of relative quantification for MMP-1 measured by real-time PCR and by ethidium bromide stained-agarose gel electrophoresis after end-point PCR. RESULTS: There was significant but very weak correlation between real-time PCR and end-point PCR for relative quantification of MMP-1 (r=0.16, P<0.01). CONCLUSIONS: Our results suggest that the use of the gel electrophoresis-based end-point PCR is inappropriate for quantifying mRNA. Therefore, in order to confirm the result of relative quantification by end-point PCR, the newly established real-time PCR method or northern hybridization should be applied.

A Simplified Approach for Detecting Homologous Deletion of SMN1 Genes in Spinal Muacular Atrophy / 医学研究杂志

Objective To develop a rapid,reliable and convenient approach for diagnosing the homozygous deletion of SMN1 gene.Methods SMN1 gene was amplified specifically with double allele-specific PCR(AS-PCR).Meanwhile,one inrelevant gene was amplified as internal control by PAGE and agarose gel electrophoresis analysis to determine whether the sick children were with homozygous deletion of SMN1 genes.Results The homozygous deletion of exon7 in SMN1 gene was identified by agarose gel electrophoresis or PAGE accurately.Conclusion Compared to PCR-RFLP and DHPLC used in the past,this approach can diagnose homozygous deletion of SMA much more accurate,easier and more convenient without completed following analyses.

IDENTIFICATION OF GLYCOSAMINOGLYCAN FROM THE BAY SCALLOP ARGOPECTEN IRRADIAUS BY AGAROSE GEL ELEC-TROPHORESIS / 中国海洋药物

Fraction F3-3-3 of glycosaminoglycan was isolated from the leftover bits of Bay scallop Argopecten irradiaus . Infrared spectra of F3-3-3 was similar to that of heparin. Agarose gel electrophoresis of this fraction was performed in the system of barbital buffer (0. 06 mol/L,pH 8. 6). It was found that F3-3-3 showed single band in the electrophoresis. Compared with the agarose gel electrophoresis of standard glycosaminoglycan, such as heparin, hyaluronic acid and chondroitin 6-sulfate, it indicated that the relative migration rate of the fraction was close to that of heparin.

Stellera chamaejasme induced apoptosis of HL-60 cells and regulated expression of bcl-2 protein in SGC-7901 cells / 中草药

Object To explore the antitumor mechanism of Stellera chamaejasme Linn. (SC). Methods SC containing-serum (SCCS) was derived from mice pretreated with different doses of SC. Cultured human leukemia HL-60 and human gastric adenocarcinoma SGC-7901 cells were used. Inhibition of proliferation was measured using MTT assay. Morphological assessment of apoptosis was performed with fluorescence microscope. DNA fragmentation was assessed by agarose gel electrophoresis and flow cytometry. Expression of bcl-2 protein was measured with immunohistochemistry. Results Exposure of exponentially growing HL-60 cells to mice serum containing 10% SC (pretreated with SC 3, 6, and 12 g/kg) for 48 h resulted in growth inhibition in a dose-dependent manner. Typical morphological changes of apoptosis and DNA fragmentation in HL-60 cells were induced. "Apobodies" in the apoptotic cells were observed, "ladder" pattern of agarose gel electrophoresis of DNA from these cells was revealed, and the percentage of apoptotic cells with fractional DNA content increased from 11.7% to 57.4%. Treatment with SC containing serum decreased the percentage of SGC-7901 cells of bcl-2 protein positive expression from 78.3% to 32.9%. Conclusion SC could induce apoptosis of HL-60 cells and decrease the expression of bcl-2 protein of gastric adenocarcinoma SGC-7901 cells.