Drug resistance analysis of Enterobacter cloacae isolated from blood samples in Hebei, 2016-2021 / 中国热带医学

@# Objective　To investigate the drug resistance of Enterobacter cloacae isolated from blood samples in 75 member units of the Bacterial Drug Resistance Monitoring Network in Hebei, 2016- 2021, so as to provide a basis for rational drug use in clinic. Methods　WHONET 5.6 software was used to retrospectively analyze drug susceptibility of Enterobacter cloacae isolated from 32 secondary hospitals and 43 tertiary hospitals. SPSS19.0 software was used for statistical analysis. Results After removing the duplicate strains, 1 225 strains of E. cloacae were isolated from blood samples of 75 hospitals during 6 years, including 157 strains from secondary hospitals and 1 068 strains from tertiary hospitals. In this study, the resistance of Enterobacter cloacae to 16 kinds of antibiotics was analyzed. The drug resistance rates to cefuroxime (52.4%-67.8%), piperacillin (27.4%-31.2%), ceftazidime (27.8%-35.5%), ceftriaxone (29.5%-45.0%), aztreonam (22.2%-32.3%), cotrimoxazole (21.6%-28.7%) were higher; the resistance rates to amikacin and tobramycin were lower than 15.0%. The resistance rates to imipenem and meropenem were 3.6%-12.3% and 5.1%-11.4%, respectively. The resistance rate to ciprofloxacin in tertiary hospitals was 22.4%, and the resistance rate to cotrimoxazole was 23.9%. Except for these two antimicrobials, the resistance rates to other antimicrobial drugs in tertiary hospitals were higher than that in secondary hospitals. A total of 121 carbapenem-resistant Enterobacter cloacae strains were detected in the past 6 years, with an increasing detection rate (χ2trend=6.305, P=0.012). Conclusions　Enterobacter cloacae has great differences in antimicrobial resistance to different antibiotics, and is sensitive to carbapenems. The drug resistance in tertiary hospitals is generally higher than that in secondary hospitals. Drug resistance monitoring and drug resistance mechanism research should be strengthened to better guide clinical drug use and curb the rise of drug resistance.

DETERMINATION OF HEMATOLOGICAL VALUES OF COMMON CROCODILE (Caiman crocodilus fuscus) IN CAPTIVITY IN THE MAGDALENA MEDIO OF COLOMBIA / Determinación de valores hematológicos de caimán común (Caiman crocodilus fuscus) en cautiverio en el Magdalena Medio de Colombia

ABSTRACT Caiman zoo breeding (Caiman crocodilus fuscus) has been developing with greater force in Colombia since the 90s. It is essential to evaluate the physiological ranges of the species to be able to assess those situations in which their health is threatened. The objective of the present study was to determine the typical hematological values of the Caiman (Caiman crocodilus fuscus) with the aid of the microhematocrit, the cyanmethemoglobin technique, and a hematological analyzer. The blood samples were taken from 120 young animals of both sexes in good health apparently (males 44 and females 76). The samples were taken from the coccygeal vein between the second and third interscalar space with lilac lid vacutainer tube whit acid ethylene-diamine-tetra-acetic as an anticoagulant. The animals are located in the middle of the Magdalena area in Colombia. The results obtained on average were Hematocrit: 23.36 %, Hemoglobin: 7.78 g/dl, Red blood cells: 3.89 106/µL, Average corpuscular volume: 60 fl, Average corpuscular hemoglobin concentration: 33.33 g/dl and Average corpuscular hemoglobin: 73.9 pg. This is the first study of hematological values for this particular species in Colombia.

RESUMEN La zoocría de babilla (Caiman crocodrilus fuscus) se ha venido desarrollando con mayor fuerza en Colombia desde los años 90. Es muy importante evaluar los rangos fisiológicos de la especie para poder valorar aquellas situaciones en las que su salud se vea comprometida. El objetivo del presente trabajo fue determinar los valores hematológicos normales del caimán común (Caimán crocodrilus fuscus) con la ayuda del microhematocrito, la técnica de cianometahemoglobina y un analizador hematológico. Las muestras de sangre se obtuvieron de 120 animales en aparente buen estado de salud, machos (44) y hembras (76); se tomaron de la vena coccígea entre el segundo y tercer espacio interescamal con tubo vacutainer tapa lila, con ácido etilen-diamino-tetra-acético como anticoagulante. Los animales están ubicados en la zona del Magdalena medio en Colombia, todos en cautiverio. Los resultados obtenidos en promedio fueron: Hematocrito: 23,36 %, Hemoglobina: 7,78 g/dl, Glóbulos rojos: 3,89 106/µL, Volumen corpuscular medio: 60 fl, Concentración de Hemoglobina corpuscular media: 33,33 g/dl y Hemoglobina corpuscular media: 73,9 pg. Este es el primer reporte de valores hematológicos para esta especie en Colombia y Latinoamérica.

Cloning, expression & evaluation of potential immunogenic recombinant capsid premembrane protein of West Nile virus

Background & objectives: West Nile virus (WNV) is a neurotropic flavivirus that has emerged globally as a significant cause of viral encephalitis. The early confirmatory diagnosis of WNV infections is important for timely clinical management and in areas where multiple flaviviruses are endemic. Diagnosis of WNV infection is primarily based on serodiagnosis, followed by virus isolation and identification. The aim of this study was to develop and evaluate a highly sensitive and specific immunoglobulin M (IgM) ELISA using the recombinant CprM protein (rWNV-CprM) for rapid, early and accurate diagnosis of WNV. Methods: The gene coding for the CprM protein of WNV was cloned and expressed in pET 28a vector followed by purification. An indirect IgM microplate ELISA using purified rWNV-CprM protein was optimized having no cross-reactivity with healthy human serum and serum samples obtained from patients with dengue and Japanese encephalitis viruses infection. Results: The comparative evaluation of this rWNV-CprM protein-specific IgM ELISA with plaque reduction neutralization test using 105 blood samples collected from patients suspected to have acute WNV infection revealed 98 per cent concordance with sensitivity and specificity of 100 and 97 per cent, respectively. Interpretation & conclusions: The recombinant CprM protein-based WNV-specific ELISA reported in this study may be useful for rapid screening of large numbers of blood samples in endemic areas during outbreaks.

Meta-analysis of effects of blood withdrawal from intravenous catheters on blood samples / 中华护理杂志

Objective To evaluate the effects of blood withdrawal from peripheral intravenous catheters (PIC) on blood samples via meta-analysis.Methods We performed a systematic search in Cochrane Library,PubMed,EBSCO,CINAHL,Embase,CMBdisc (China Biology Medicine disc),Wan Fang Data,VIP (China Science and Technology Journal Database),CNKI (China National Knowledge Infrastructure)to collect randomized controlled trails (RCTs)about the effects of blood withdrawal from peripheral intravenous catheters.The quality of RCTs was critically appraised and data were extracted by two reviewers independently.Meta-analysis was conducted for eligible RCTs.Results Totally 22 studies were retrieved and 7554 blood samples were included.Results of meta-analysis showed as compared with standard blood drawn by direct venipuncture (DVP),there was no significant risk of hemolysis rates of blood samples collected from PVC [random effect RR =2.01,95％ CI (1.01,4.00),P=0.05];with certain procedures,results of laboratory tests of blood samples withdrawal from PIC had little impacts on red blood cell count(RBC),white blood cell count(WBC),platelet count (PLT),fasting blood-glucose(FBG)and serum levels of sodium,potassium,calcium,chlorine,and the differences had no statistical significance (P＞0.05).Conclusion Blood samples withdrawal from PIC will not increase hemolysis if sampling after infusion has been stopped for 2～3 min and 2～5 ml blood is discharged,which are reliable for blood routine and biochemical tests.After a comprehensive consideration,the technique of blood withdrawal from a PIC could be used interchangeably from DVP.

Analysis of the significance of whole blood sample in the diagnosis of severe cases of hand, foot and mouth disease / 中国生化药物杂志

Objective To study the diagnostic significance of whole blood sample index in the severity of hand, foot and mouth disease, and to analyze its pathogenesis. Methods 194 cases of severe cases of hand, foot and mouth disease were selected as the case group,and 193 cases of mild hand, foot and mouth disease were selected as the control group. Blood samples were collected after admission to study the whole blood samples, including white blood cells, erythrocytes, platelets, medium cell ratio, lymphocyte ratio, mononuclear cell ratio, C-reactive protein test; The patient's immune system, the nervous system were also tested. Results Compared with patients with mild group, the white blood cell level of the patients in the severe group was higher and the albumin level was higher, the difference was statistically significant (P<0.05), the monocyte ratio, the T cell ratio was lower, the difference was statistically significant (P<0.05). Conclusion The white blood cell level and monocyte ratio of patients with severe hand, foot and mouth disease have changed. The whole blood sample has a certain sensitivity to the diagnosis of severe hand, foot and mouth disease.

Rapid Surface-Enhanced Raman Spectroscopic Detection of Blood Cyanide Based on Pinhole Shell-Isolated Nanoparticleand Online Lysis-Purging and Trapping Approach / 分析化学

A rapid and simple method for the determination of cyanide in blood was developed based on pinhole shell-isolated nanoparticles (pinSHINs)-enhanced Raman spectroscopy and an online lysis-purging and trapping approach.In the online lysis-purging and trapping device, the bound cyanide in blood can be cleaved through sulfuric acid acidification, and transferred into HCN volatile gas, then purged into alkaline solution to form NaCN solution, thus high-efficient liberation and entrapment of cyanide from the methemoglobin-bound form can be achieved.The pinSHINs substrate is quite stable to weaken the gold-dissolution effect caused by cyanide under strong alkaline condition, and therefore the detection window can be prolonged to 1 h comparing with 5 min of AuNPs.A limit of detection down to 10 μg/L and a linear range from 100-2000 μg/L in blood were achieved in this method.This method was further applied to rapid measurement of blood samples of cyanide exposed rats and clinic poisoned patients, which provided a sensitive, selective and reliable way for rapid detection of cyanide poisoning.

Detection of Circulating Tumor Cells in Breast Cancer Patients Using Cytokeratin-19 Real-Time RT-PCR

PURPOSE: The roles of circulating tumor cells (CTCs) as predictive and prognostic factors, as well as key mediators in the metastatic cascade, have been investigated. This study aimed to validate a method to quantify CTCs in peripheral blood using a real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay for cytokeratin (CK)-19 and to evaluate the utility of this assay in detecting CTCs in breast cancer patients. MATERIALS AND METHODS: Real-time monitoring PCR of fluorescently labeled specific hybridization probes for CK-19 mRNA was established. Peripheral blood samples from 30 healthy donors, 69 patients with early breast cancer, 47 patients with locally advanced breast cancer, and 126 patients with metastatic breast cancer were prospectively obtained and analyzed for CTC detection. RESULTS: CK-19 mRNA was not detectable in healthy subjects using the real-time RT-PCR method. The detection rates of CK-19 mRNA in breast cancer patients were 47.8% for early breast cancer (33/69), 46.8% for locally advanced breast cancer (22/47), and 61.1% for metastatic breast cancer (77/129). The detection rate of CK-19-positive CTCs in metastatic disease was slightly higher than early or locally advanced breast cancer; however, the detection rate according to disease burden was not statistically different (p=0.097). The detection rate was higher in patients with pleural metastasis (p=0.045). CTC detection was associated with poor survival (p=0.014). CONCLUSION: A highly specific and sensitive CK-19 mRNA-based method to detect CTCs in peripheral blood in breast cancer patients can be used in further prospective studies to evaluate the predictive and prognostic importance of CTCs.

Analysis of the pre-analytical phase in a private pathology laboratory of Maringá city-PR, Brazil / Análise da fase pré-analítica em laboratório privado da cidade de Maringá-PR, Brasil

ABSTRACT Introduction: The pre-analytical phase involves most of the errors of clinical analysis laboratories. Information characterizing the occurrence of these errors can be useful to provide prevention and reduction strategies. Objective: The aim of this study was to identify the main errors in the pre-analytical phase of a private laboratory in the city of Maringa-PR, and their occurrence rate. Materials and methods: Data were collected from sample recollection orders from June 2013 to May 2015. A total of 707,449 exams collections were registered, and 1,274 (0.18%) of these had ordered a new collection based on some criteria adopted by the laboratory. Conclusion: The pre-analytical phase represented 70.8% of the orders for new collection. According to the reports from the Quality Control Database, the most frequent reason of recollection in the preanalytical phase was insufficient sample volume (58.54%), followed by clotted sample (19.29%), hemolyzed sample (9.43%), incorrect labeling of sample (8.21%), lipemic sample (3%) and improper sample (1.56%). Although the results of nonconformity forms presented different records, both clearly point to problems in collection and patient registration sector, sustaining that the main problems of clinical laboratories arise from the failure in quality specifications for pre-analytical phase.

RESUMO Introdução: A fase pré-analítica envolve a maior parte dos erros dos laboratórios de análises clínicas. Informações que caracterizem a ocorrência desses erros podem ser úteis para disponibilizar estratégias de prevenção e redução. Objetivo: O objetivo principal deste estudo foi identificar os principais erros na fase pré-analítica de um laboratório privado da cidade de Maringá-PR e sua frequência de ocorrência. Materiais e métodos: Foram coletados dados dos pedidos de recoleta de junho de 2013 a maio de 2015. Ao todo, foram cadastradas coletas de 707.449 exames; destes, 1.274 (0,18%) tiveram pedidos de nova coleta com base em algum critério adotado pelo laboratório. Conclusão: A etapa pré-analítica representou 70,80% dos pedidos de nova coleta. Segundo os relatórios do Banco de Dados do Controle de Qualidade, o motivo mais frequente de recoletas na etapa pré-analítica foi material insuficiente (58,54%), seguido por amostra coagulada (19,29%), amostra hemolisada (9,43%), identificação errada (8,21%), amostra lipêmica (3%) e material errado (1,56%). Embora os resultados dos formulários de não conformidades apresentassem registros diferentes destes, ambos apontam claramente problemas no setor de coleta e cadastro do paciente, confirmando que os principais problemas dos laboratórios clínicos provêm da falta de especificações de qualidade para a fase pré-analítica.

Application of dilution regression method in the coagulation detection of fat blood samples / 国际检验医学杂志

Objective To explore the dilution regression method of coagulation detection(PT ,APTT) in fat blood samples . Methods We collected 40 normal blood coagulation specimens (no fat blood ,no jaundice ,no hemolysis) in Yan′an hospital of Kun-ming ,then we detected the PT and APTT of the original plasma and 3-fold diluted plasma and 5-fold diluted plasma ,the we used the data both of before dilution and diluted to do the linear regression analysis ,and finally we got the regression equations of each index .we also collected 33 fat blood samples in Yanan hospital of Kunming ,which be divide into three groups through the severity of triglycerides :mild fat blood group(1 .7 mmol/L≤TG0 .05) .Conclusion Dilution regression method can be used to detect the fat blood samples in the clinical coagulation detection .

Changing malaria status among children in Ngapyawtone village, Letpandan Township.

Peripheral blood samples were collected from 60 children (2 - 9 years) residing in Ngapyawtone village, Yayngan village tract, Gway Dauk Kwin Rural Health Centre, Letpandan Township, Bago Division in March, 2012. Temperature, anaemia, jaundice, spleen and liver enlargement were assessed for clinical parameters associated with malaria. Parasite positivity by malaria microscopy and detection of malaria antigen by rapid diagnostic test were done. The malaria parasite prevalence by blood slide microscopy was 8.3%. The malaria parasite prevalence by RDT was also 8.3%. Rapid Immunochromatographic test was performed for Pf/Pv Antibodies. Malaria Antibody positivity rate was 5 % in the study population. Changing pattern of malaria (parasite prevalence, serological prevalence and spleen rate) among children due to effective malaria control program was found in Ngapyawtone village where similar study was done in 1995.

Staphylococcus aureus DNA in human venous blood detected by real-time quantitative PCR assay / 中华临床营养杂志

Objective To establish a rapid real-time quantitative polymerase chain reaction (RQ-PCR)assay in quantifying and detecting Staphylococcus aureus DNA from human venous blood samples,so as to quantificationally evaluate the systemic infection caused or deteriorated by intestinal bacteria translocation.Methods Totally 26 clinical blood samples and 15 simulation blood samples were detected.The primers and TaqMan probe were designed targeting the highly conserved house-keeping femA gene of Staphylococcus aureus,and a 20 μl RQ-PCR amplification reaction system was established.The standard curve was built based on the recombinant plasmid DNA containing the amplicon of the target gene,and genomic DNA was extracted using QIAamp DNA Blood Mini Kit.Results The specificity of primers and probe was excellent,the detecting limit was 100 copies/μl (103 CFU/ml),the sensitivity was 99.7％,and the specificity was 94.6％.The correlation coefficient of the standard curve was between 0.9918 and 0.9997.For samples with different Staphylococcus anreus concentrations,the average accuracy of the RQ-PCR assay was (96.25 ± 2.26) ％ ; the intra-and interassay coefficients of variation were (8.06 ±0.07)％ and (10.01 ±4.40)％,respectively.The average recovery rate of Staphylococcus aureus DNA in blood samples was (111.72 ± 20.72) ％.In clinical blood samples,the positive rate of Staphylococcus aureus DNA was 15.4％ (4/26),while the blood culture of these samples all produced negative result for Staphylococcus aureus.Conclusion RQ-PCR assays is a rapid,sensitive,and specific method with good repetitiveness and can be used in the quantitative detection of Staphylococcus aureus in whole blood samples.

Avaliação do hiperinsulinismo em amostra de crianças pré-puberes / Hyperinsulinism assessment in a sample of prepubescent children

OBJETIVOS: Determinar a relação entre o índice de massa corporal (IMC), o homeostasis model assessment - insulin resistance (HOMA-IR) e a insulinemia. MÉTODOS: Realizou-se um estudo observacional prospectivo transversal com 132 crianças pré-púberes em idade escolar e residentes no município de Santo André (SP). Fez-se a avaliação antropométrica e a mensuração da glicemia, da insulinemia e do índice HOMA-IR. RESULTADOS: Dentre as 132 crianças avaliadas, 78 eram meninas (59,1 por cento) e 54 eram meninos (40,9 por cento), com média de idade de 8,7 anos e média de IMC de 13,7 kg/m². Observou-se uma associação significativa e positiva entre HOMA-IR e IMC, insulina e IMC, peso e HOMA e entre insulina e peso; também foi constatado que, quanto maior for o IMC, maior será o valor de HOMA. CONCLUSÕES: Os resultados do presente estudo permitem concluir que há uma forte associação entre o hiperinsulinismo e a obesidade, devendo ser tomadas algumas medidas para evitar o ganho de peso durante a infância e a adolescência.

OBJECTIVES: To determine the relationship between body mass index (BMI), homeostasis model assessment - insulin resistance (HOMA-IR) and insulinemia. METHODS: This was a prospective cross-sectional observational study of 132 prepubescent schoolchildren residents in the municipality of Santo André, Brazil. Children underwent anthropometric assessment, their glycemia and insulinemia were measured and their HOMA-IR index calculated. RESULTS: Seventy-eight of the 132 children (59.1 percent) were girls and 54 were boys (40.9 percent), with a mean age of 8.7 years and mean BMI of 13.7 kg/m². A significant positive association was detected between HOMA-IR and BMI, insulin and BMI, weight and HOMA and between insulin and weight. It was also found that the higher the BMI, the greater the HOMA score. CONCLUSIONS: The results of this study allow for the conclusion that there is a strong association between hyperinsulinism and obesity. Measures should be taken to avoid weight gain during childhood and adolescence.

Comparison of outcomes and efficiency between drawings of two blood samples and three blood samples for culture.

Health economic analysis is one of the strategies for healthcare provider or policy makers to make decision. Cost identification is the basic evaluation while cost minimization or cost benefit is a further step comparing the cost with the efficacious outcome. Ideally, the comparison between drawings of two blood samples and three blood samples for culture has to be done by prospective randomization to avoid selective biases as much as possible. In the situation of a cohort study like “Comparison of Outcomes and Efficiency between Drawings of Two Blood Samples and Three Blood Samples for Culture,” the reasons for taking two or three blood samples have to be clearly stated. Also, all the demographical data of subjects such as gender, age group, volume of each blood sample, duration between blood drawing and diagnosis have to be comparable and stated as much as possible. First blood specimens from table 1 and table 2 show the recovery rate of positive hemoculture which are summarized as follows. Although we could draw a conclusion that each subject has 11-12% chance to have positive first blood culture specimen, but drawing of two blood specimens were more contaminated (20.1%) than three blood specimens (8.3%). The evidence is clear and comparable. Regarding the calculation of Number Needed to Treat (NNT) of 56 cases or doing the third hemoculture will find one more case than only drawing two specimens, we had 10,264 specimens/year of two specimens and would miss 184 cases/that could be detected by the third hemoculture and just only one resulted this true infection. How many subjects responsible for the cost of over treat or under treat? How much is the cost of life and reputation of the hospital? Interpretation and conclusion in terms of monetary unit provide ultimate outcome as morbidity mortality.

Integrons and Multi-drug Resistance among Pseudomonas aeruginosa Strains Isolated from Blood Samples / 中华医院感染学杂志

OBJECTIVE To investigate the incidence of integrons in Pseudomonas aeruginosa strains isolated from blood samples in Peking University People′s Hospital and to analyze the correlation between integrons and drug resistance of P.aeruginosa.METHODS Forty-two strains of clinically isolated P.aeruginosa were collected.The antibiotics susceptibility was tested by K-B methods.Integrase gene of integron was amplified by PCR using degenerate primers.The integrons were classified by restriction fragment length polymorphism(RFLP) analysis of positive PCR products with Hinf Ⅰ restriction enzyme.RESULTS The drug-resistance rates of 42 strains of P.aeruginosa against 20 kinds of antibiotics ranged from 9.5% to 100%.Twenty-three strains were resistant to 12 kinds of antibiotics.Nineteen of the 42 isolates(45.2%) contained integrons,all of which were revealed as class Ⅰ of integrons by RFLP analysis.Neither class Ⅱ nor class Ⅲ of integron was detected.The positive percentage of integrons was increased by years.CONCLUSIONS Class Ⅰ integrons are widespread in isolates from blood samples in our hospital.The presence of integrons is closely associated with multi-drug resistance of P.aeruginosa.

Detection of filarial infection using Wuchereria bancrofti microfilariae culture antigen and filter paper blood samples in enzyme linked immunosorbent assay.

Blood collected on filter paper by finger-prick gave results comparable to intravenous serum samples when analysed by enzyme-linked immunosorbent assay (ELISA). All the 100 microfilaraemia, 5 out of 100 endemic normals and none of the 10 nonendemic normal filter paper blood samples showed the presence of filarial antibody when tested by this method,using culture antigen and anti-immunoglubulins, class G, Μ and A — penicillinase conjugate. When the same samples were screened for the presence of IgM antibody, 91 out of 100 microfilaraemia, 13 out of 100 endemic normal and none of the 10 nonendemic normal samples showed a positive reaction. Enzyme linked immunosorbent assay, using culture antigen and filter paper blood samples, appears to work in large field studies for detection of filarial infection.