Ontogeny of digestive enzymes in larvae of the Clown Anemonefish, Amphiprion ocellaris (Perciformes: Pomacentridae)

Introduction: The Clown anemonefish (Amphiprion ocellaris) is the most popular fish species in the marine aquarium trade; however, there is a lack of information on their digestive physiology during larval ontogeny, valuable information needed for diet design and management protocols. Objective: To characterize the early digestive enzymes of A. ocellaris larvae. Methods: We used three pools (10 larvae each) and extracted 10 samples per tank, from just before hatching to the 38th day. We analyzed the specific activity of acid and alkaline proteases, trypsin, chymotrypsin, leucine aminopeptidase and lipase; and did acid and alkaline protease zymograms. Results: We detected all measured enzymes at hatching. Acid proteases increased in activity until the 38th day. Alkaline proteases, trypsin, chymotrypsin, and leucine aminopeptidase had the same pattern, and maximum activity on the 8th day, decreasing at the 38th day. Lipase activity peaked on the 8th and 30th day. The acid zymogram had a single band, appearing on the 8th day. A total of eight alkaline proteases were revealed (154.2, 128.1, 104.0, 59.8, 53.5, 41.9, 36.5 and 25.1 KDa), with seven bands on the 1st day and all bands from the 3rd to 8th day, decreasing at two bands (41.9 and 25.1 KDa) in the 38th day. Conclusion: A. ocellaris has a functional stomach on the 8th day, and, on the 38th day, a digestive omnivore pattern with a tendency to carnivory.

Introducción: El pez payaso (Amphiprion ocellaris) es la especie de pez más popular en el comercio de acuarios marinos; sin embargo, falta información sobre su fisiología digestiva durante la ontogenia larval, información valiosa necesaria para protocolos de diseño y manejo dietético. Objetivo: Caracterizar las enzimas digestivas tempranas de larvas de A. ocellaris. Métodos: Usamos tres homogenados (con 10 larvas cada uno) y extrajimos 10 muestras por tanque, justo antes de la eclosión hasta el día 38. Analizamos la actividad específica de proteasas ácidas y alcalinas, tripsina, quimotripsina, leucina aminopeptidasa y lipasa; e hicimos zimogramas de proteasas ácidas y alcalinas. Resultados: Detectamos todas las enzimas medidas en la eclosión. La actividad de proteasas ácidas incrementó hasta el día 38. Proteasas alcalinas, tripsina, quimotripsina, y leucina aminopeptidasa tuvieron el mismo patrón, con actividad máxima en el octavo día, decreciendo en el día 38. Hubo picos en la actividad lipasa a los ocho y 30 días. El zimograma ácido tuvo una banda única, apareciendo al octavo día. Se hallaron ocho proteasas alcalinas (154.2, 128.1, 104.0, 59.8, 53.5, 41.9, 36.5 y 25.1 KDa), con siete bandas al primer día, y todas las bandas entre el tercer y octavo día, bajando a dos bandas (41.9 y 25.1 KDa) al día 38. Conclusión: A. ocellaris tiene un estómago funcional al octavo día, y, al día 38, un patrón digestivo omnívoro con tendencias carnívoras.

Determination of bioavailable fluoride in toothpastes marketed in Medellín, Colombia, through capillary electrophoresis / Fluoruro biodisponible en cremas dentales comercializadas en Medellín ­ Colombia mediante electroforesis capilar

Currently, the FDI recommendation (World Dental Federation) for fluoride concen­tration in toothpastes is that the total fluoride content declared by manufacturers be between 1,000 and 1,500 ppm, and of this amount, at least 800 ppm should be bioavai­lable fluoride. The aim of this study was to describe the market of toothpastes marketed in Medellín and to measure the concentration of bioavailable fluoride by capillary elec­trophoresis to verify their compliance with the FDI recommendation for the prevention of dental caries. After sampling the toothpastes available for sale in the 16 communes of the city of Medellin, a single operator measured in triplicate the concentration of bioavailable fluoride in 36 samples of previously blinded toothpastes using the capi­llary electrophoresis (EC) technique. Most of the evaluated toothpastes that declared a content of 1,000­1,500 ppm fluoride met the recommendation of presenting at least 800 ppm bioavailable fluoride. Measurement of bioavailable fluoride in toothpastes with MFP (sodium monofluorophosphate) is recommended, as it is known that binding to the abrasive can decrease its concentration over time.

Actualmente la recomendación de la FDI (Federación Dental Internacional) sobre la concentración de fluoruro en cremas dentales es que el contenido de fluoruro total de­clarado por los fabricantes sea entre 1.000 y 1.500 ppm, y que de esta cantidad al menos 800 ppm sea fluoruro biodisponible. El objetivo de este estudio fue describir el mercado de las cremas dentales comercializadas en Medellín y medir en estas la concentración de fluoruro biodisponible por electroforesis capilar para verificar su cumplimiento de la recomendación de la FDI para la prevención de la caries dental. Luego de realizar un mues­treo en las 16 comunas de la ciudad de Medellín sobre las cremas dentales disponibles a la venta, un solo operador midió por triplicado la concentración de fluoruro biodisponible en 36 muestras de cremas dentales previamente cegadas, mediante la técnica de electroforesis capilar (EC). La mayoría de las cremas dentales evaluadas que declaraban contenido de 1.000 a 1.500 ppm de fluoruro, cumplieron la recomendación de presentar al menos 800 ppm de fluoruro biodisponible. Se recomienda realizar la medición del fluoruro biodisponible en las cremas con MFP, ya que se conoce que la unión al abrasivo puede disminuir su concentración en el tiempo.

Differences in the distribution of hemoglobin variants according to the geographic regions in a Colombian population

ABSTRACT Introduction: Colombia has been subject to intense genetic and cultural currents due to its geographical location. Hemoglobinopathies are the most common recessive diseases found worldwide and represent an important public health problem, according to the region and ancestry of each country. Objectives: To evaluate the frequency of hemoglobin variants according to the geographical region in a population group adjusted to sex and age in Colombia. Methods: This was a descriptive retrospective study of hemoglobin variants performed by electrophoresis in patients treated at and/or referred to specialized care institutions in Bogota, Colombia between January 2009 and December 2020. Results: A total of 2,224 results were analyzed, 48.4% male and 51.5% female; 63.3% of patients were without alterations, 14.3% presented with thalassemia, 17.3%, HbS, 2.3%, HbS/C, 1.8%, HbC, 0.5%, HbE and 0.5% persistent HbF, with HbS being more prevalent in males (p = 0.005). When assessing the geographical regions of Colombia, a higher prevalence of HbS was found in the Pacific (p = 0.005) and Caribbean regions, while Thalassemia and HbS were more prevalent in the Andean and Orinoquia regions, and it was rare to find any hemoglobinopathies (p = 0.0001) in the Amazonian region. Conclusions: The main hemoglobinopathies found in Colombia are HbS, predominantly in males, and Thalassemia. The distribution of hemoglobinopathies in different geographical regions of Colombia is influenced by ancestry.

Albumin-to-protein ratio in spot urine samples for analysis of proteinuria selectivity in chronic kidney disease / Razão albumina/proteína em amostras isoladas de urina para análise da seletividade de proteinúria na doença renal crônica

ABSTRACT Introduction: The albumin-to-creatinine ratio and total protein-to-creatinine ratio in spot urine samples have already been validated as surrogates for 24-hour albuminuria and proteinuria measurements. Thus, we hypothesized that the type of proteinuria, detected by the electrophoretic pattern of 24-hour urine, could be predicted by the simple proportion of albumin in the total urine protein content, using the albumin-to-protein ratio (APR). Our study sought to validate the use of APR as a cheaper substitute for urinary protein electrophoresis (UPE). Methods: Using different mathematical models, we compared, the albumin fraction in 24-hour urine samples by electrophoresis and the APR ratio in spot samples from 42 outpatients with chronic kidney disease (CKD). Results: A strong log-order correlation r = 0.84 (0.75-0.92; 95% CI, p = 0.001) was observed between APR and the albumin fraction in the UPE. Conclusion: The APR can substitute electrophoresis in CKD outpatients.

Resumo Introdução: A utilização da razão albumina/creatinina e da razão proteína total/creatinina em amostras isoladas de urina já foram validadas como substitutos para a albuminúria e proteinúria em 24 horas. Assim, nossa hipótese é que o tipo de proteinúria, dado pelo padrão eletroforético da urina de 24 horas, poderia ser previsto pela simples proporção de albumina no conteúdo total de proteínas na urina, utilizando a razão albumina/proteína (RAP). O presente estudo procurou validar o uso da RAP como um substituto mais prático e de menor custo da eletroforese de proteínas urinárias (EPU). Métodos: Foram utilizados diferentes modelos matemáticos a fim de comparar a fração de albumina pela eletroforese em amostras de urina de 24 horas e a RAP em amostras isoladas em 42 pacientes ambulatoriais com doença renal crônica. Resultados: Foi observada uma forte correlação logarítmica r = 0,84 (0,75-0,92; 95% CI, p = 0,001) entre a RAP e a fração de albumina pela EPU. Conclusão: A RAP pode substituir a eletroforese urinária em pacientes renais crônicos ambulatoriais.

Simultaneous detection of thirteen exons of dystrophin gene by optimized multiplex PCR assay to screen Duchenne/Becker muscular dystrophy

Advancements in Polymerase Chain Reaction (PCR) technology and other techniques like Deoxyribonucleic acid (DNA)signal and target amplification have become key procedures in molecular diagnostics. PCR enables the synthesis of nucleic acids in vitro through which a DNA segment can be specifically replicated in a semiconservative way that sets forth deletion and mutation analysis. Multiplex PCR (M-PCR) is beneficial over standard and long PCR as this can amplify more than one locus using the respective primer sets. In harmony with this, the present study aimed to optimize M-PCR followed by its chemistry and condition to screen Duchenne Muscular Dystrophy (DMD) [OMIM #310200] and Becker Muscular Dystrophy (BMD) [OMIM #300376]. Muscular Dystrophies (MDs) are a broad group of hereditary, progressive, and degenerative disorders of muscles. X-linked recessive D/BMD are caused by mutation/s in the dystrophin gene [OMIM #300377] that encodes for dystrophin protein [UniProt#P11532]. As dystrophin is the human metagene with 79 exons, mutational analysis is very challenging. Chamberlain set (10 plex), Beggs set (9 Plex), and Kunkel set (7 Plex) is used for many years to diagnose this condition. However, in this study, Beggs set is customized with 13 exons to screen DMD gene mutation in a single reaction. Optimization of M-PCR was designed with many physicochemical parameters. According to the literature and after many appraisals the present study demonstrated the most sufficient concentration of various chemical components and optimal cycling conditions to optimize the modified Beggs set (13 Plex). 50 µL PCR reaction includes primer(s) (0.3–0.5 µM each), dNTP mixture (160 µM each), Dream Taq buffer (1X), Taq DNA polymerase (6U/50 µL), DNA template (250 ng/50 µL), BSA (0.4 µg/µL), and MgCl2 (1.4 mM). To get the most effective results cyclic conditions obtained were 10 min initial denaturation at 94°C, 62°C annealing temperature, and 35 PCR cycles at 72°C extending temperature. Consequently, the study successfully formulated a less expensive and simple approach for >3000 bp that was used to screen D/BMD. Finally, a developed M-PCR mix with a unique combination of specificity and sensitivity coupled with great flexibility has led to a true revolution in molecular diagnostics.

Functional and Hemodynamic Efficacy of Non-Poliferative Diabetic Retinopathy Treatment by Endonasal Electrophoresis of Tanakan

Aims: To study the effectiveness of the treatment of patients with non-proliferative diabetic retinopathy by tanakan endonasal electrophoresis according to functional and hemodynamic data.Study Design: Cross-sectional comparative analysis.Place and Duration of Study: Department of Ophthalmology, clinic of Tashkent Medical Academy, between 2017 and 2020.Methodology: We included 66 patients (123 eyes), 23 men and 43 women; age range 18-75 years with non-proliferative diabetic retinopathy. The patients were divided into 2 groups: to receive daily tablets of Tanakan (control), or daily endonasal electrophoresis of Tanakan (main) within ten days. Before and after the course of therapy, the patients underwent determination of visual acuity, Doppler ultrasound imaging of the eye and computer static perimetry.Results: Improvements in visual acuity were observed in 87,3% of the main group patients, and in 22% of the control group. Statistically significant increase in blood flow velocity and a decrease in the resistance index were observed in the main group (P < 0.05). Retinal photosensitivity increased by 22% in the main group, and by 10% in the control group. The mean deviation from the age norm decreased by 33% in the main group and by 12% in the control group. Among the patients of the main group, 30% experienced a decrease in absolute scotomas and 100% in relative scotomas. Among the patients of the control group, 21% and 83% experienced a decrease in absolute and relative scotomas, respectively.Conclusion: Treatment with tanakan endonasal electrophoresis significantly improved visual acuity, eye hemodynamics, and retinal photosensitivity. This treatment is more effective than the traditional use of ginkgo- biloba tablets.

Microbial detection and analysis of a foodborne disease caused by Staphylococcus aureus infection / 中国基层医药

Objective:To identify the cause of a suspected foodborne disease caused by bacterial food poisoning using multiple detection methods.Methods:At 9:35 a.m. on August 9, 2022, a suspected foodborne disease incident was handled by the Hefei Center for Disease Control and Prevention. The preliminary laboratory test results showed that the foodborne disease was caused by Staphylococcus aureus infection. Sixteen strains of Staphylococcus aureus were isolated, cultured, and identified using real-time fluorescent polymerase chain reaction. At the same time, the 16 strains of Staphylococcus aureus isolated from different sources were analyzed by pulsed-field gel electrophoresis to determine the correlation between strains. Results:The distribution of 16 strains of Staphylococcus aureus enterotoxins isolated was that the anal swab and chopping board (inner) smear samples from a salesperson surnamed Li showed SEB type, and the remaining 14 strains all carried type A, C, and E enterotoxin genes simultaneously. Pulsed-field gel electrophoresis typing results showed that there were two types of strains; except the anal swab and chopping board (inner) smear samples from the salesperson surnamed Li, the other 14 strains were all of the same type, with a similarity of 100%. The similarity between the anal swab and the chopping board (inner) smear samples from the salesperson surnamed Li was 80.65%, and the similarity with another type was 59.26%. This result was consistent with the detection results of Staphylococcus aureus enterotoxins. Conclusion:The foodborne disease in this case was caused by mixed contamination of two or three different sources of Staphylococcus aureus with enterotoxins.

Analysis of serum protein electrophoresis results in patients with acute brucellosis / 中华地方病学杂志

Objective:To understand the results of serum protein electrophoresis in patients with acute brucellosis, and to provide objective indicators for clinical diagnosis and treatment of acute brucellosis patients.Methods:Thirty acute brucellosis patients diagnosed by the Center for Disease Prevention and Control of Donghe District, Baotou City from January to June 2021 were selected as the case group, and 30 local healthy persons who took physical examination during the same period were selected as the control group. Blood samples were collected from the two groups of people, serum was separated and serum protein electrophoresis was performed. Independent sample t-tests were used to compare total protein concentrations, protein fractions (albumin, α1-globulin, α2-globulin, β-globulin and γ-globulin) and albumin/globulin (A/G) ratio. Results:The age of patients in the case group was (52.35 ± 6.74) years old, with 25 males and 5 females. The course of the disease ranged from 0.3 to 2.6 months, with an average of 1.78 months. The age of people in the control group was (52.74 ± 5.47) years old, with 25 males and 5 females. There were no statistically significant difference in age and sex distuibution between the two groups ( P > 0.05). The concentrations of total protein and α2-globulin fractions were not significantly different between the two groups ( t = 0.78, 2.04, P > 0.05, respectively). Compared with the control group, the differences in the percentage of albumin, α1-globulin, β-globulin, γ-globulin in total protein and A/G ratio were statistically significant ( t = 4.78, 4.76, 2.89, 2.20, 3.65, P < 0.05). Conclusions:The percentage of serum albumin in total protein and A/G ratio decrease in patients with acute brucellosis, while the percentage of α1-globulin, β-globulin and γ-globulin in total protein increase. The percentage of albumin, α1-globulin, β-globulin, γ-globulin in total protein and A/G ratio can be uesed as objective indicators for clinical treatment of acute brucellosis.

Recent applications and chiral separation development based on stationary phases in open tubular capillary electrochromatography(2019-2022) / 药物分析学报

Capillary electrochromatography(CEC)plays a significant role in chiral separation via the double sepa-ration principle,partition coefficient difference between the two phases,and electroosmotic flow-driven separation.Given the distinct properties of the inner wall stationary phase(SP),the separation ability of each SP differs from one another.Particularly,it provides large room for promising applications of open tubular capillary electrochromatography(OT-CEC).We divided the OT-CEC SPs developed over the past four years into six types:ionic liquids,nanoparticle materials,microporous materials,biomaterials,non-nanopolymers,and others,to mainly introduce their characteristics in chiral drug separation.There also added a few classic SPs that occurred within ten years as supplements to enrich the features of each SP.Additionally,we discuss their applications in metabolomics,food,cosmetics,environment,and biology as analytes in addition to chiral drugs.OT-CEC plays an increasingly significant role in chiral separation and may promote the development of capillary electrophoresis(CE)combined with other instruments in recent years,such as CE with mass spectrometry(CE/MS)and CE with ultraviolet light detector(CE/UV).

Etiologic identification and drug susceptibility analysis of a Citrobacter freundii food poisoning event / 中国热带医学

@#Abstract: Objective　In order to provide reference for emergency treatment of a sudden food poisoning incident, pathogen detection and drug resistance analysis were carried out. Methods　Diarrheal stool and surplus food samples were detected by GB 4789 and the isolates were identified by VITEK2 and matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), at the same time, the bacterial drug sensitivity test was carried out by using the method of microbroth dilution, and the isolates from different sources were molecularly classified by pulsed field gel electrophoresis (PFGE), and the correlation between the strains was analyzed by BioNumerics software. Results　Totaly 13 leftovers and 3 diarrhea patients were isolated and identified, The total number of colonies and coliforms in 7 leftovers samples all exceeded the standard, and Citrobacter freundii was detected in 5 leftovers and 2 stools. The results of drug sensitivity test showed that seven strains of Citrobacter freundii were sensitive to ciprofloxacin, tetracycline, chloramphenicol, gentamicin, amikacin, cefotaxime and meropenem, but completely resistant to ampicillin, and there was no multiple drug resistance. The results of pulsed field gel electrophoresis (PFGE) showed that 7 strains of Citrobacter freundii had the same PFGE bands and 100% homology, showing the same clone. Conclusions　This food poisoning incident was caused by Citrobacter freundii. The pathogen of food poisoning can be quickly and accurately determined by MALDI-TOF MS, which is beneficial to the early diagnosis and treatment of infectious diseases. It is suggested to strengthen the corresponding management, improve food safety awareness and prevent similar incidents.

Contamination status and molecular typing of Legionella pneumophila in artificial water environment in Shanghai hospitals from 2019 to 2020 / 环境与职业医学

Background The incidence of Legionnaires' disease is increasing globally and artificial water environment is becoming a common source of outbreaks. Molecular typing techniques can help prevent and control Legionella. Objective To understand the molecular epidemiological characteristics of Legionella pneumophila in artificial water environment of Shanghai hospitals, and provide a scientific basis for the prevention and control of Legionnaires' disease. Methods Water samples were collected from artificial water environment in 14 hospitals from May to October each year from 2019 to 2020 in Shanghai. A total of 984 water samples were collected from 8 Grade-A tertiary hospitals and 6 non-Grade-A tertiary hospitals, including 312 samples of cooling water, 72 samples of chilled water, and 600 samples of tap water. The water samples were isolated and serotyped for Legionella pneumophila and preserved, and the positive rate of Legionella pneumophila in the samples was used as an indicator of contamination. The preserved strains were resuscitated and 81 surviving strains were obtained for pulsed field gel electrophoresis (PFGE) typing analysis. Results A total of 124 Legionella pneumophila positive water samples were detected, with a positive rate of 12.60%. The positive rate was higher in the Grade-A tertiary hospitals (16.54%, 87/526) than in the non-Grade-A tertiary hospitals (8.08%, 37/458) (χ2=15.91, P<0.001). The positive rate of cooling water (23.40%) was the highest among different types of water samples, and the difference was statistically significant (χ2=61.19, P<0.001). The difference in positive rate of tap water was statistically significant among different hospital departments (χ2=11.37, P<0.05). The positive rate in 2019 (15.06%) was higher than that in 2020 (9.84%) (χ2=6.23, P<0.05). From May to October, August had the highest annual average positive rate (16.46%) and October had the lowest (8.54%), but the difference in positive rates among months was not statistically significant (χ2=5.39, P=0.37). The difference in positive rate among districts was statistically significant (χ2=24.88, P<0.001). A total of 131 strains of Legionella pneumophila were isolated, with serotype 1 (80.15%, 105/131) predominating. Among the 81 surviving strains of Legionella pneumophila subjected to PFGE typing, the band-based similarity coefficients ranged from 41.30% to 100%. Among the 29 PFGE band types (S1-S29) recorded, each band type included 1-10 strains, and S28 was the dominant band type. Four clusters (I-IV) of PFGE band types were identified, accounting for 66.67% (54/81) of all strains and containing 13 band types. Conclusion Legionella pneumophila contamination is present in the artificial water environment of hospitals in Shanghai from 2019 to 2020, and the contamination in tap water deserves attention. The detected serotype of Legionella pneumophila is predominantly type 1, and PFGE typing reveals the presence of genetic polymorphism. Therefore, the monitoring and control of Legionella pneumophila in hospital artificial water environment should be strengthened.

Etiological characteristics of Vibrio parahaemolyticus strains isolated from foodborne outbreaks in Zhongshan in 2019 - 2021 / 公共卫生与预防医学

Objective To investigate the etiological characteristics of food poisoning isolates of Vibrio parahaemolyticus (VP) from 2019 to 2021 in Zhongshan City. Methods A total of 37 strains of Vibrio parahaemolyticus isolated from 8 food poisoning incidents in Zhongshan City from 2019 to 2021 were collected, including 1 residual food isolate and 36 human isolates. The genetic correlation of Vibrio parahaemolyticus food poisoning isolates in this region was analyzed by serological typing, virulence gene detection (TLH, TDH, and TRH), drug sensitivity test, pulsed field gel electrophoresis (PFGE) and multipoint sequence typing (MLST). Results The 37 strains of Vibrio parahaemolyticus were divided into 4 serotypes: O3:K6, O10:K4, O4:K8, and O4:KUT. The tdh+ and trh- were the main virulence genotypes, accounting for 97.30% (36/37). The drug resistance rate of cefazolin was 40.54% (15 strains R, 22 strains I), and no multidrug-resistant strains were found. The 37 VP strains were divided into 23 PFGE types and 6 cluster groups, with correlation coefficients ranging from 60.4%-100%. The multipoint sequencing typing showed that the 37 VP strains were divided into 9 ST types and 3 complex groups, of which ST3 type was the main type (23 strains, 62.1%). Conclusion This study has found that the dominant virulence types of Vibrio parahaemolyticus food poisoning isolates in Zhongshan City from 2019 to 2021 are tdh+ and trh-, and 37 representative strains can be divided into 6 PFGE clusters and 9 ST types with MLST type being mainly ST3. This study has identified the rare serotype O10:K4 which has caused an increase in the proportion of food poisoning events, suggesting that we should strengthen detection and be alert to the risk of continued local epidemics of new rare serotype strains.

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.

Anti-Alzheimer's Disease Effect of Polysaccharides from Zanthoxyli Pericarpium and Analysis of Its Structural Characteristics by Sugar Spectrum / 中国实验方剂学杂志

ObjectiveThe therapeutic effect of polysaccharides from Zanthoxyli Pericarpium on Alzheimer's disease（AD） was evaluated through establishing a mouse model of AD， and the structural characteristics of the polysaccharides was analyzed by sugar spectrum. MethodThe AD model of mice with rapid aging was established by intraperitoneal injection of D-galactose combined with gavage of aluminum trichloride， and the learning and memory ability of mice was evaluated by Morris water maze test， the histopathological status of brain and neuronal damage were observed by hematoxylin-eosin（HE） staining and Nissl staining. After hydrolysis of polysaccharides from Zanthoxyli Pericarpium with acid and different glycosidases， the characteristics of hydrolysates were analyzed by high performance thin layer chromatography（HPTLC） and fluorescence assisted carbohydrate gel electrophoresis（PACE）. HPTLC chromatography was performed on a silica gel 60 plate with sampling volume of 5 μL， developing solvent of ethyl acetate-glacial acetic acid-water（2∶2∶1）， developing twice， aniline-diphenylamine-phosphoric acid solution as chromogenic agent， and heating at 105 ℃ for 10 min， and then observed under sunlight. PACE experimental conditions were 34% separation gel and 8% concentration gel， electrophoresis buffer was 0.1 mol·L-1 tris（hydroxymethyl） aminomethane（Tris）-boric acid buffer（pH 8.2）. Electrophoresis was carried out at 0 ℃ and the loading amount was 3-6 μL. The sample ran to the front of the gel with a constant current of 15 mA， and imaged under ultraviolet 365 nm. ResultThe results of Morris water maze test showed that polysaccharides from Zanthoxyli Pericarpium significantly improved the learning and memory ability of AD model mice， shortened the escape latency， and significantly increased the number of crossing and the residence time in the target quadrant. The results of histopathological experiments showed that polysaccharides from Zanthoxyli Pericarpium could improve the pathological conditions and neuronal damage in the CA1 and CA3 regions of hippocampus of AD mice， and the number of Nissl corpuscles was significantly increased. The results of sugar spectrum analysis showed that the results of HPTLC and PACE analysis were basically consistent， polysaccharides from Zanthoxyli Pericarpium could be mainly hydrolyzed into small molecular sugars by cellulase and pectinase， indicating that they mainly contained β-1，4-glucosidic bond and α-1，4-galacturonic acid glycosidic bond， and could be slightly hydrolyzed by glucanase， β-galactosidase and β-mannase， indicating that they contained only a small amount of α-1，6-glucosidic bond， β-galactosidic bond， β-1，4-mannosidic bond. ConclusionPolysaccharides from Zanthoxyli Pericarpium has obvious therapeutic effect on AD mice， and its structure mainly contains β-1，4-glucosidic bond and α-1，4-galacturonic acid glycosidic bond， which can provide a reference for the structural analysis of traditional Chinese medicine polysaccharides.

Causes of Abnormal Hemoglobin Electrophoresis / 中国实验血液学杂志

OBJECTIVE@#To investigate the possible causes of abnormal hemoglobin electrophoresis results.@*METHODS@#The hemoglobin electrophoresis results of 5 696 patients in the First Affiliated Hospital of Chengdu Medical College from September 2018 to July 2021 were collected, and the abnormal results and clinical significance were analyzed.@*RESULTS@#The results of 486 patients (accounting for 8.53%) were abnormal, of which 300 cases had increased HbA2, 135 cases had decreased HbA2, 44 cases had increased F alone, and 7 cases had abnormal hemoglobin bands. Among the 486 patients, 246 patients were thalassemia gene positive (the positive rate was 50.62%), including 29 cases of α thalassemia, 208 cases of β thalassemia and 9 cases of αβ thalassemia. Among the patients with elevated HbA2, 68.67% were detected β thalassemia, 3.00% αβ thalassemia, 9.33% were suspected to be caused by macrocytosis, 6.33% by thyroid dysfunction, and 12.67% by uncertainty of the method. Among the patients with reduced HbA2, 21.48% were detected α thalassemia, 60.00% iron deficiency anemia, 8.15% were suspected to be caused by thyroid dysfunction, and 10.37% by uncertainty of the method. Among the patients with elevated F alone, the results of thalassemia gene detection were negative, 40.91% of them were suspected to be caused by macrocytosis, 27.27% by hereditary persistence of fetal hemoglobin, 29.55% by special physiological condition of pregnant women, and 2.27% by hyperthyroidism. Abnormal hemoglobin bands were detected in 7 patients, including 4 cases of hemoglobin D, 2 cases of hemoglobin E, and 1 case of hemoglobin J.@*CONCLUSION@#Thalassemia, iron deficiency anemia, macrocytosis such as megaloblastic anemia and non-severe aplastic anemia, thyroid dysfunction, hereditary persistence of fetal hemoglobin, abnormal hemoglobin diseases, the uncertainty of the method are all important causes of abnormal hemoglobin electrophoresis results. In clinical work, the patient's indicators should be comprehensively analyzed to determine the possible cause.

Development and clinical application of hydrogel formulations containing papain and urea for wound healing

Abstract Hydrogels are used for wound treatment, as they may contain one or more active components and protect the wound bed. Papain is one of the active substances that have been used with this purpose, alongside urea. In this paper, carboxypolymethylene hydrogels containing papain (2% and 10% concentrations) and urea (5% concentration) were produced. Physical-chemical stability was performed at 0, 7, 15 and 30 days at 2-8ºC, 25ºC and 40ºC, as well as the rheological aspects and proteolytic activity of papain by gel electrophoresis. Clinical efficacy of the formulations in patients with lower limb ulcers was also evaluated in a prospective, single-center, randomized, double-blind and comparative clinical trial. The results showed 7-day stability for the formulations under 25ºC, in addition to approximately 100% and 15% of protein activity for 10% and 2% papain hydrogel, respectively. The rheological profile was non-Newtonian for the 10% papain hydrogel tested. There were no significant differences regarding the mean time for healing of the lesions, although 10% papain presented a better approach to be used in all types of tissue present in the wound bed.

Electroforesis de isoenzimas de fosfatasa alcalina: utilidad diagnóstica en pacientes con enfermedad de células falciformes / Alkaline phosphatase isoenzyme electrophoresis: diagnostic utility in patients with sickle cell disease

Introducción: Las fosfatasas alcalinas (FAL) humanas son indicadores enzimáticos de daño de órganos. Las complicaciones de la drepanocitosis incluyen lesión de órganos debido a la reperfusión, la vasculopatía proliferativa y la anemia hemolítica. Objetivo: Demostrar la utilidad de la determinación de la actividad enzimática de la FAL y la movilidad electroforética de las isoenzimas séricas de FAL en pacientes con drepanocitosis en estado basal y durante las crisis. Métodos: Estudio descriptivo, longitudinal, prospectivo en 85 individuos adultos, 25 controles sanos y 60 pacientes, entre enero y diciembre/2018. Variables estudiadas: genotipos (SS/Sβo, SC/Sβ+), edad, sexo, actividad de FAL e isoenzimas por electroforesis en gel de agarosa con y sin lectina: hepática 1 (H1) y (H2); placenta 1(P1), ósea, intestinales 1, 2, 3 (I1.2.3) en estado basal y crisis. Resultados: La actividad de la FAL fue significativamente mayor en los pacientes que en los controles. Hubo diferencias significativas en la actividad de la FAL y de la fracción H1+P1/1 de pacientes en estado basal en relación con el grupo control. La actividad de la enzima y las isoenzimas no mostró diferencias significativas entre los genotipos. Igual comportamiento se observó en la actividad de las enzimas e isoenzimas durante las crisis vasoclusivas dolorosas y hepáticas. Se encontraron diferencias significativas en la actividad de la fracción hepática H1+P1/1 entre los grupos de 20-29 y 40-49 años Conclusiones: La determinación de la FAL en los pacientes con drepanocitosis es útil y permite establecer un perfil isoenzimático personalizado, con importancia pronóstica como un marcador biológico de alarma(AU)

Introduction: Human alkaline phosphatases (ALP) are enzymatic indicators of organ damage. Complications of sickle cell disease include injury to organs due to reperfusion injury, proliferative vascular disease, and hemolytic anemia. Objective: To demonstrate the usefulness of determing ALP activity and the electrophoretic mobility of the serum ALP isoenzymes in patients with sickle cell disease at base line and during the crises. Methods: A descriptive, longitudinal, prospective study was carried out in 85 adult individuals, 25 healthy controls and 60 patients, between January and December/2018. Studied variables: genotypes (SS/SβO, SC/Sβ+), age, sex, ALP activity and isoenzymes by agarose gel electrophoresis with and without lectin: hepatic 1 (H1) and (H2); placenta 1 (P1), bone, intestinal 1, 2, 3 (I1.2.3) in basal state and crisis. Results: ALP activity was significantly higher in patients than in controls. Significant differences were found in the activity of the ALP and fraction H1+P1/1 of patients in basal state in relation to the control group. The activity of the enzyme and the isoenzymes showed no significant differences between genotypes. The same behavior was observed in the activity of enzymes and isoenzymes during painful and liver vasooclusive crises. Significant differences were found in the activity of the liver fraction H1+P1/1 between the groups of 20-29 and 40-49 years. Conclusions: The determination of the ALP in patients with sickle cell disease is useful and allows to establish a personalized isoenzimatic profile, with prognostic importance as a biological alarm marker(AU)

HemoTypeSC point-of-care testing shows high sensitivity with alkaline cellulose acetate hemoglobin electrophoresis for screening hemoglobin SS and SC genotypes

ABSTRACT Introduction: By providing timely actionable results for prompt management, point-of-care testing (POCT) kits have revolutionised medical care for various diseases, ranging from infectious diseases like malaria to genetic disorders, such as sickle cell disease (SCD). They are, however, underutilised in the diagnosis of SCD in developing countries, where the need is greatest. Objective: The study was aimed at assessing the sensitivity of HemoTypeSC POCT among a cohort of children with SCD, previously diagnosed by Alkaline cellulose acetate hemoglobin electrophoresis (ACAE), with or without high-performance liquid chromatography (HPLC). Methods: In this descriptive cross-sectional study, HemoTypeSC test was conducted on all participants and its sensitivity was determined by comparing results with those obtained using ACAE. Discordance was verified with HPLC. Results: One hundred and forty-five children aged one to 19 years were studied. There were 84 males and 61 females (male: female ratio = 1.4:1). The HemoTypeSC was able to correctly diagnose sickle cell anemia (SCA) and hemoglobin SC in all (100%) of the children tested. Conclusion: The HemoTypeSC shows high sensitivity in detecting SCA and hemoglobin SC. Hence, it is useful for targeted screening of individuals suspected of having SCD, leading to rapid diagnosis of these hemoglobinopathies, even in resource-constrained settings.

Experiencia en el uso de los protocolos Biomed-2 para el estudio de reordenamientos de TCR e inmunoglobulinas en proliferaciones linfoides en el Instituto Nacional de Cancerología, Colombia / Experience with the BIOMED-2 standardized polymerase chain reaction protocol for immunoglobulin and T- cell receptor clonality analysis in the Instituto Nacional de Cancerología, Colombia

Introducción. El consorcio europeo BIOMED-2 se creó para determinar si una población linfoide de difícil clasificación patológica es clonal. En Colombia, la implementación de estas pruebas comenzó en el 2015 en el Instituto Nacional de Cancerología E.S.E. (Bogotá). Objetivos. Determinar el comportamiento de las pruebas de reordenamiento clonal o clonalidad linfoide. y determinar las dificultades de su uso en nuestro medio verificando su adaptación local y los resultados en una serie retrospectiva de casos y consecutiva de proliferaciones linfoides sometidas a los protocolos BIOMED-2. Materiales y métodos. A partir de las historias clínicas, se recolectaron los datos clínicos e histológicos y los resultados de los análisis de los reordenamientos en todos los casos de proliferaciones linfoides sometidas a los protocolos BIOMED-2, entre febrero de 2015 y mayo de 2019. Resultados. Se hallaron 132 casos, de los cuales 47 se clasificaron mediante los protocolos de Biomed-2 como hiperplasias linfoides reactivas, 62 como linfomas T, 19 como linfomas B y 3 como neoplasias linfoides de linaje no establecido. Solo en un caso falló la extracción de ADN. Según estos resultados, la mayor dificultad diagnóstica para el patólogo fue el análisis de los infiltrados linfoides T, la mayoría (44) de los cuales correspondía a lesiones cutáneas. Conclusiones. Las pruebas de clonalidad pueden usarse en tejidos de diversa calidad en nuestro medio como ayuda en el diagnóstico de proliferaciones linfoides de difícil clasificación. Es importante hacerlas e interpretarlas de manera multidisciplinaria y considerar cada caso por separado.

Introduction: The European BIOMED-2 consortium was created to evaluate clonality in lymphoproliferations that are difficult to diagnose. In Colombia, the implementation of these tests began in 2015 at the Instituto Nacional de Cancerología E.S.E., Bogotá. Objectives: To determine the behavior of the rearrangement tests for lymphoid clonality and the difficulties of its implementation in our field through a series of retrospective and consecutive cases of lymphoid proliferation subjected to the BIOMED-2 protocols. Materials and methods: Clinical and histological data and the results of the rearrangement analysis of all cases of lymphoid proliferation subjected to the BIOMED-2 protocols between February 2015 and May 2019 were collected from clinical histories. Results: We recovered 132 samples from which 47 corresponded to reactive lymphoid hyperplasias, 62 to T lymphomas, 19 to B lymphomas, and three to lymphoid neoplasms of unestablished lineage. Only in one case did DNA extraction fail. According to these results, the greatest diagnostic difficulty for the pathologist was the analysis of T lymphoid infiltrates, most of which (44) were skin lesions. Conclusions: Clonality tests can be used in tissues of different quality to help in the diagnosis of lymphoid proliferations that are difficult to classify. It is important to implement and interpret them in an interdisciplinary way considering each case separately.

Valor diagnóstico de la electroforesis de proteínas y cadenas ligeras libres en suero en el mieloma múltiple / Diagnostic value of serum protein electrophoresis and free light chains in the multiple myeloma's diagnosis

RESUMEN Fundamento: La electroforesis de proteínas y las cadenas ligeras libres en suero son técnicas utilizadas en el diagnóstico del mieloma múltiple. Sin embargo, la utilidad diagnóstica de ambas pruebas puede variar según el método empleado y condiciones reales del medio donde se realicen. Objetivo: Determinar el valor diagnóstico de la electroforesis de proteínas y de las cadenas ligeras libres en suero en el mieloma múltiple. Metodología: Se realizó un estudio retrospectivo de los parámetros electroforesis de proteínas en suero y cadenas ligeras libres en suero a 43 pacientes con diagnóstico de mieloma múltiple por evaluación de la médula ósea. La electroforesis de proteínas se realizó por el método convencional de separación de proteínas sobre papel de acetato de celulosa y para las cadenas ligeras libres se aplicó un ensayo inmunoturbidimétrico en el que se usó un analizador químico (Cobas 311). Se calcularon 7 parámetros que evaluaron la exactitud diagnóstica. Resultados: Todos los parámetros que evaluaron la exactitud diagnóstica estuvieron dentro de los intervalos de confianza en ambas pruebas. Conclusiones: La electroforesis de proteínas y las cadenas ligeras libres en suero son ensayos de gran utilidad en el diagnóstico del mieloma múltiple y se deben utilizar en conjunto para la mayor captación posible de casos.

ABSTRACT Background: Protein electrophoresis and serum free light chains are techniques used in the diagnosis of multiple myeloma. However, the diagnostic utility of both tests may vary according to the method used and the actual conditions of the environment where they are performed. Objective: To determine the diagnostic value of protein electrophoresis and serum free light chains in multiple myeloma. Methodology: A retrospective study of serum protein electrophoresis parameters and serum free light chains was conducted in 43 patients diagnosed with multiple myeloma by bone marrow evaluation. Protein electrophoresis was completed by the conventional method of protein separation on cellulose acetate paper and for free light chains an immunoturbidimetric assay was applied in which a chemical analyzer (Cobas 311) was used. Seven parameters were calculated to evaluate diagnostic accuracy. Results: All parameters assessing diagnostic accuracy were within confidence intervals in both tests. Conclusions: Protein electrophoresis and serum free light chains are very useful assays in the diagnosis of multiple myeloma and should be used in conjunction for the highest possible approval of cases.