Dried blood spots: a robust tool for malaria surveillance in countries targeting elimination

With the advancements in analytical and molecular techniques, Dried Blood Spots (DBS) are re-emerging as attractive and cost-effective alternatives for global health surveillance. The use of DBS has been well-characterized in the neonatal screening of metabolic diseases, therapeutic screening as well as in epidemiological studies for biomonitoring. Malaria is one such infectious disease where DBS use can expedite molecular surveillance for assessing drug resistance and for refining drug usage policies. In India, malaria cases have reduced significantly over the past decade but to achieve malaria elimination by 2030, country-wide DBS-based screening should be conducted to identify the presence of molecular markers of artemisinin resistance and to study parasite reservoirs in asymptomatic populations. DBS has wide applications in genomics, proteomics, and metabolomic studies concerning both host and pathogen factors. Hence, it is a comprehensive tool for malaria surveillance that can capture both host and parasite information. In this review, we elucidate the current and prospective role of DBS in malaria surveillance and its applications in studies ranging from genetic epidemiology, parasite and vector surveillance, drug development and polymorphisms to ultimately how they can pave the roadmap for countries aiming malaria elimination

Case reports of two children with neurofibromatosis type 1 / 中国热带医学

@#Abstract: Objective　To report the clinical characteristics and genetic test results of two children with neurofibromatosis type 1 (NF1), and to provide reference for the comprehensive diagnosis-treatment and follow-up plans of NF1 patients based on the existing diagnosis and treatment progress of NF1. Methods　Two children with NF1 admitted to the Department of Children's Medicine, Haikou people's Hospital in May and June 2022 were selected to analyze the clinical data of their clinical manifestations, laboratory examination, genetic test results, diagnosis and treatment and follow-up retrospectively. Results　Two children had typical clinical manifestations, such as café-au-lait spots, axillary freckles, intraocular iris hamartoma. Venous blood was collected from case 1 and his parents for NF gene test, and a new mutation of c.4084C>T in the NF1 gene was found, and their parents did not have the pathogenic gene; the venous blood of the children in case 2 was tested for whole-exome gene analysis, and a heterozygous nonsense variant c.910C>T:p.R304 on the NF1 gene was found, , which was verified by Sanger sequencing to be inherited from his mother, his mother has café-au-lait spots and brain glioma, and has undergone surgery to remove the brain glioma, but has not undergone chemoradiotherapy or targeted therapy. No neurological malignancies were detected in either of the two children at follow-up until July 2022. Conclusions　The clinical manifestations of NF1 are relatively typical, genetic testing is conducive to determine its classification, and regular follow-up review can help to detect and treat malignant tumors early, thus improving the patient's quality of life.

Analysis of hot spots and trends of the standardized residency training of Traditional Chinese Medicine based on the CiteSpace knowledge map software / 中华医学教育探索杂志

Objective:To analyze the research progress, hot spots and future trends of standardized residency training of Traditional Chinese Medicine from 1998 to now by using CiteSpace knowledge map software.Methods:The related literature on standardized residency training of Traditional Chinese Medicine were retrieved on CNKI. And after Refworks format conversion, using CiteSpace 5.6 R3 software for the construction of knowledge map, the author cooperation, institutional cooperation, keyword co-occurrence, and time-sharing research hot spots views of related literature on standardized residency training of Traditional Chinese Medicine were drawn respectively.Results:A total of 259 papers were obtained, with 253 authors. Ren Xianqing and Cui Jin published the most articles (each with 4 papers), and Beijing University of Chinese Medicine published the most articles (with 23 papers). There were 259 keywords in total, and 16 keywords with frequency greater than 5 and centrality greater than 0.1. There were 14 keyword cluster tags, and 15 burst terms were detected in total, and the trend discussion was carried out accordingly.Conclusion:The research trend of the standardized residency training of Traditional Chinese Medicine mainly follows the relevant policies issued by the functional departments of the government, and the research hot spots depend on the obstacles and difficulties encountered in the practical application of each training base.

Chromatic disorder in Trachops cirrhosus (Spix, 1823) (Chiroptera: Phyllotomidae) on Ecuadorian Amazon

We present a case of chromatic disorder observed in a specimen of Trachops cirrhosus from lowland forests of the Ecuadorian Amazon. During the field work, six individuals of this species were captured with mist nets in an intervened habitat, of which, a juvenile male specimen was distinguishable from the rest of the individuals due to the presence of whitish spots on the ventral region, on the chest and abdomen. We discuss the previous reports of this type of pigmentary disorders in bats from Ecuador, and in the Neotropic, documenting the first record of Trachops cirrhosus with this condition for the country.

Presentamos un caso de desorden cromático observado en un ejemplar de Trachops cirrhosus proveniente de bosques de tierras bajas de la Amazonía ecuatoriana. Durante el trabajo de campo, seis individuos de esta especie fueron capturados con redes de niebla en un hábitat intervenido, de los cuales, un ejemplar macho juvenil fue distinguible del resto de individuos debido a la presencia manchas blanquecinas en la región ventral, en el pecho y abdomen. Discutimos sobre los reportes previos de este tipo de trastornos pigmentarios en murciélagos de Ecuador, y en el Neotrópico, documentando el primer registro de esta especie con esta condición para el país.

A Cross Sectional Study On The Prevalence Of Vitamin A Deficiency

Background: Vitamin A is an essential nutrient needed for normal vision, immune functions, maintenance of epithelial cell integrity, growth and development, reproductive function. It is the most important cause of preventable childhood blindness in developing countries. To estimate the prevalence of vitamin A deficiency in children ?15 yearsAim: reporting to a tertiary care centre A cross sectional study was conducted on 100 children agedMaterials & Methods: upto 15years reporting to eye OPD from April 2022- June 2022. Detailed history from mother was collected. A comprehensive ophthalmic examination including visual acuity measurement, anterior segment, fundus evaluation was done. Vitamin A deficiency was clinically assessed by using WHO classification of Xerophthalmia. TheResults: prevalence of clinical vitamin A deficiency in this study was 9%. Maximum presentation was seen in age group of >2-5 years. Prevention, early recognition by regular screening and nutritional education would decrease theConclusion: risk of developing clinical vitamin A deficiency

Concentración espacial de las denuncias por violencia familiar en Ciudad Victoria: un acercamiento mediante la detección de hot spots / Spatial concentration of complaints of family violence in Ciudad Victoria: an approach through the detection of hot spots

Resumen La violencia familiar es el principal delito cometido a nivel nacional en México. Tamaulipas tiene una tasa de denuncias que se encuentra por encima de la media nacional desde el año 2016 hasta el 2018, destacando el municipio de Vitoria con la tasa más alta en ese período. El objetivo de este trabajo fue identificar la concentración espacial de las denuncias por violencia familiar en Ciudad Victoria y especificar la relación entre las desventajas del vecindario y la ubicación en un hot spot (punto caliente). El estudio se basa en la teoría de la desorganización social y tiene como soporte los datos de la Fiscalía General del Estado de Tamaulipas en el periodo 2016-2018. Para ello, se desarrolló el análisis exploratorio de datos espaciales mediante las técnicas de estimación de densidad de kernel, promedio del vecino más cercano, análisis de hot spot Gi* de Getis-Ord y un análisis confirmatorio mediante una regresión logística binaria, procesamientos desarrollados en el software ArcMap 10.8.1 y en el paquete estadístico R. Los hallazgos indicaron que las denuncias se producen desde un número reducido de unidades geográficas. La prueba del promedio del vecino más cercano mostró que existe agrupamiento estadísticamente significativo (z = - 10.825, P = 0.000). En el análisis Gi* de Getis-Ord se detectó que 602 manzanas (9.8 % del total) son hot spots (al 95 % y 99 % de confianza) de alta incidencia, mientras que la regresión logística confirmó que la cantidad de ocupantes por vivienda y los hogares con jefatura femenina están asociados positivamente con la probabilidad de estar dentro de un hot spot de violencia familiar. El patrón de denuncias mostró poca dispersión geográfica y asociación estadística relevante con las variables de desorganización social.

Abstract The main crime committed in Mexico is domestic violence. Tamaulipas had a reporting rate above the national average from 2016 to 2018, with Ciudad Victoria having the highest rate during that period. The objective of this work is to identify the spatial concentration of complaints on family violence and to specify the relationship between the neighborhood characteristics and its location in a hot spot. To that aim, the study uses the social disorganization theory and data of complaints taken from the Fiscalia General del Estado de Tamaulipas during the 2016-2018 period. The exploratory analysis of spatial data was developed by using kernel density estimation techniques, nearest neighbor average, Getis-Ord Gi* hot spot analysis and confirmatory analysis through binary logistic regression, with the help of the ArcMap 10.8.1 software and R statistical package. The findings indicated that complaints of domestic violence are produced from a small number of geographic units. The nearest neighbor mean test showed that there is a statistically significant grouping of complaints (z = - 10.825, P = 0.000). In the Gi * analysis of Getis-Ord, 602 blocks (9.8 % of the total) were identified as hot spots (at 95 % and 99 % confidence), while the logistic regression confirmed that the number of occupants per dwelling and households headed by women are positively associated with the probability of being in a hot spot of family violence. The pattern of the complaints showed little geographic dispersion and relevant statistical associations with the social disorganization variables.

McCune-Albright syndrome - A case report with transmission electron microscopy

Abstract McCune - Albright syndrome is a genetic disease with cutaneous mosaicism caused by post-zygotic activating mutations in GNAS locus, it has a triad of fibrous bone dysplasia, café-au-lait macules and precocious puberty. We examined a 22-year-old female patient with café au lait spot in right side of the abdomen, with a chessboard - like distribution, extending to right thigh with geographical contours, she has also an ovarian cyst, scoliosis and truncal obesity. Biopsies were taken from the hyperpigmented area and processed for light microscopy and for transmission electron microscopy. Light microscopy showed increased melanin pigment with HE staining. Immunohistochemistry with melanocytic markers (HMB-45 and Melan-A) revealed a normal number of melanocytes. Transmission electron microscopy demonstrated normal epidermal structures, such as desmosomes, cytokeratin filaments and hemidesmosomes. With high magnifications an irregular melanossomal contour was seen, with some indentations in their outline.

Study on the inhibition and remineralization of enamel demineralization by polydopamine in vitro / 口腔疾病防治

Objective@#To investigate the inhibitory effect of polydopamine (PDA) on enamel demineralization in isolated teeth and the induction of hydroxyapatite (HA) production on the surface of demineralized enamel to provide a novel protocol for the prevention and treatment of enamel demineralization. @*Methods@#Twenty isolated bovine teeth were cut into 20 enamel slices and randomly divided into an experimental group and a control group, with 10 slices in each group. The enamel slices in the experimental group were immersed in 2 mg/mL freshly prepared dopamine solution and incubated for 24 hours at room temperature in the dark to prepare the PDA coating, while the control group was left untreated. Then, the isolated bovine teeth, with and without PDA coating, were immersed in artificial demineralization solution at 37 °C for 3 days, followed by 7 days in simulated body fluid (SBF), and the immersion solution was changed daily. The surface morphology of enamel was observed by scanning electron microscopy (SEM), the calcium/phosphorus ratio of the enamel surface was analyzed by energy dispersive spectroscopy (EDS), and the characteristic functional groups in enamel deposits were analyzed by Fourier transform infrared spectroscopy (FTIR).@* Results@#Compared with the control group, the number of demineralized pores produced after 3 d of enamel demineralization with polydopamine coating was less, and the diameter was smaller. EDS elemental analysis showed that the Ca/P ratio after enamel demineralization was 2.37 in the experimental group, which was smaller than the 2.53 ratio in the control group. In the remineralization experiment, after 7 days of remineralization of PDA coated enamel in the experimental group, lamellar grains were produced on the enamel surface, and the growth showed obvious directionality, growth regularity and uniform arrangement. In the control group, the surface of enamel was flocculent mineral deposit, and the crystallinity was poor. The FTIR results proved that the enamel surface deposit of PDA-coated enamel was HA after 7 d of remineralization. @*Conclusion @#PDA can affect the nucleation process of HA and promote the production of HA on the surface of demineralized enamel.

Estudo de estabilidade de anfetaminas e produtos de biotransformação de cocaína e tetraidrocanabinol em amostras de urina seca em papel (dried urine spots) / Stability study of amphetamines, biotransformation products of cocaine and tetrahydrocannabinol in dried urine spots

O número de pessoas utilizando substâncias ilícitas de forma recreativa aumenta a cada ano, chamando a atenção de estudiosos de diversas áreas do conhecimento. Com isso, a demanda de exames toxicológicos exigida para trabalhadores, vítimas de crimes e esportistas também tem crescido. A amostra biológica mais utilizada para análises toxicológicas continua sendo a urina, visto que sua obtenção é menos invasiva, possibilita coletar grande volume de amostra e pode-se detectar substâncias até dias após ter ocorrido a exposição ou consumo. Entretanto, estas amostras necessitam de um grande volume físico para serem armazenadas e transportadas aos laboratórios, devendo ser mantidas em temperatura baixa e controlada para conservação. Outro ponto a se considerar é a quantidade de amostra insuficientemente coletada, ou extravasamento do conteúdo, contaminando outras amostras e muitas vezes, inviabilizando a análise. Uma alternativa recente para tais problemas é utilizar a técnica chamada de dried urine spots (DUS), onde poucos microlitros de urina são colocados em um papel absorvente e secos sob temperatura ambiente, preservando de agentes degradantes os componentes presentes na urina. Assim, o objetivo deste trabalho é avaliar a estabilidade das substâncias do presente estudo em alta temperatura, temperatura ambiente e em temperaturas de 4°C e -20°C. Para este fim, foi necessário desenvolver, validar e aplicar métodos de extração e determinação de anfetaminas e produtos de biotransformação de cocaína e tetraidrocanabinol carboxílico (THCCOOH) em amostras dried urine spot, utilizando cromatografia líquida acoplada à espectrometria de massas. Os picos foram identificados por UPLC-ESI-MS/MS, com tempo total de 5 mins utilizando fase A- água, formiato de amônio e 0,1% ácido fórmico, e B- metanol: acetonitrila (6:4) + 0,1% de ácido fórmico. A extração foi feita utilizando acetonitrila: metanol: acetona (1:1:1) +ácido fórmico 0,1%. Não foi possível iniciar a validação de THCCOOH, visto uma possível complexação do analito com o papel. Para as outras substâncias, o método cromatográfico desenvolvido se mostrou eficiente e seletivo, com LOD e LOQ de 10 ng/mL para todos os analitos, sendo linear até 1000 ng/mL, atendeu as especificações de precisão e exatidão e carryover. As amostras permaneceram estáveis ao longo de 32 dias nas temperaturas estudadas, demonstrando a segurança em se utilizar a técnica de DUS para armazenamento e transporte de amostras biológicas dentro da faixa de temperatura do estudo até 32 dias

The number of people using illegal substances in a recreational way increases each year, drawing the attention of scholars from different areas of knowledge. As a result, the demand for workplaces drug tests, toxicological tests for victims of crimes and dopping has also grown. The biological sample most used for toxicological tests remains urine, since obtaining it is less invasive, it is possible to collect a large volume of sample and it is possible to detect substances up to days after exposure or consumption has occurred. However, these samples require a large physical volume to be stored and transported to the laboratories, and must be kept at a low temperature for conservation. Another point to consider is the amount of sample insufficiently collected, or leakage of the content, causing contamination of other samples and often making the analysis unfeasible. A recent alternative to such problems is to use "dried urine spots" (DUS), where few microliters of urine are placed on absorbent paper and dried at room temperature, preserving the components present in the urine from degrading agents. Thus, the objective of this work is to evaluate the stability of the substances in this study at high temperature, room temperature and at temperatures of 4°C and -20°C. For this purpose, it was necessary to develop, validate and apply methods of extraction and determination of amphetamines and biotransformation products of cocaine and carboxylic tetrahydrocannabinol (THCCOOH) in dried urine spot samples, using liquid chromatography coupled to mass spectrometry (LC-MS). The peaks were identified liquid chromatography coupled to a mass spectrometer (UPLC-ESI-MS/MS), with a total time of 5 mins using phase A- water, ammonium formate and 0.1% formic acid, and B- methanol: acetonitrile (6:4) + 0.1% formic acid. Extraction was done using acetonitrile: methanol: acetone (1:1:1) + 0.1% formic acid. It was not possible to perform the validation of THCCOOH, given a possible complexation of the analyte with the paper. To the others substances, the chromatographic method developed proved to be efficient and selective, with LOD and LOQ of 10 ng/mL for all analytes, being linear up to 1000 ng/mL, meeting the specifications of precision and accuracy and carryover. The samples remained stable for 32 days at the temperatures studied, demonstrating the safety of using the DUS technique for storage and transport of biological samples until 32 days on temperature range studied

A escuta psicanalítica do traumático social ­ ênfase racismo / Psychoanalytic listening of social trauma ­ emphasis on racism

Neste trabalho a palavra equidade vai ressoar em direção ao funcionamento das instituições, em particular as de psicanálise, e à escuta analítica do traumático social, com ênfase no racismo. As instituições tendem a ser conservadoras em sua natureza, mas são compostas por pessoas que se renovam no decorrer do tempo e podem abrir-se para mudanças. Ao se tornarem permeáveis aos sofrimentos e mal-estar que advém do campo social e comunitário podem adotar posicionamentos que favoreçam seu envolvimento nessas problemáticas. É destacado como a escuta analítica pode encontrar pontos cegos quando os/as analistas/terapeutas se fecham em bolhas de ressonância mútua, não se sensibilizando e nem se colocando na pele do outro diferente de si mesmo ou de seu grupo social. Torna-se fundamental perceber como a cultura racista onde fomos criados impregna nosso inconsciente, através dos Ideais do Eu, com a crença de que o branco seria a medida do que é universal e belo e, ao mesmo tempo, negando nossa realidade racista.(AU)

In this paper the word equity will resonate towards the functioning of institutions, in particular those of psychoanalysis, and the listening to the social trauma, with an emphasis on racism. Institutions tend to be conservative in nature, but they are made of people who renew themselves over time and can be open to change. By becoming permeable to the sufferings that come from the social and community field they may adopt positions that favor their envolvement in these issues. It highlights that analytical listening can find blind spots when analysts/therapists close themselves in bubbles of mutual resonance, not putting themselves in the shoes (skin) of others, diferents from themselves or their social group. It becomes fundamental to understand how the racist culture where we were raised impregnates our unconscious, through the ideals of the Ego, with the belief that white people would be the measure of what is universal and beautiful and, at the same time, denying our racist reality.

En este trabajo la palabra equidad resonará hacia el funcionamiento de las instituciones, en particular las del psicoanálisis, y a la escucha analítica del trauma social, com énfasis en el racismo. Las instituciones tienden a ser de naturaleza conservadora, pero están formadas por personas que se renuevan con el tiempo y pueden abrirse a cambios. Al volverse permeable a los sufrimientos y malestares que vienen del campo social y de la comunidad pueden adoptar posiciones que favorezcan su implicación en estos problemas. Se destaca que la escucha analítica puede encontrar puntos ciegos cuando los analistas/terapeutas se cierran en burbujas de resonancia mutua, no sensibilizarse y no ponerse en la piel de otra persona que no sea uno mismo o su próprio grupo social. Se hace imprescindible comprender cómo la cultura racista en la que nos criamos impregna nuestro inconsciente, a través de los Ideales del Yo, con la creencia de que el blanco sería la medida de lo universal y bello y, al mismo tiempo, negar nuestra realidade racista.(En este trabajo la palabra equidad resonará hacia el funcionamiento de las instituciones, en particular las del psicoanálisis, y a la escucha analítica del trauma social, com énfasis en el racismo. Las instituciones tienden a ser de naturaleza conservadora, pero están formadas por personas que se renuevan con el tiempo y pueden abrirse a cambios. Al volverse permeable a los sufrimientos y malestares que vienen del campo social y de la comunidad pueden adoptar posiciones que favorezcan su implicación en estos problemas. Se destaca que la escucha analítica puede encontrar puntos ciegos cuando los analistas/terapeutas se cierran en burbujas de resonancia mutua, no sensibilizarse y no ponerse en la piel de otra persona que no sea uno mismo o su próprio grupo social. Se hace imprescindible comprender cómo la cultura racista en la que nos criamos impregna nuestro inconsciente, a través de los Ideales del Yo, con la creencia de que el blanco sería la medida de lo universal y bello y, al mismo tiempo, negar nuestra realidade racista.(AU)

Standardization of an organic DNA extraction method from dried blood spots and its downstream molecular applications in neonatal screening and diagnostic confirmation of lysosomal disorders

Introduction: Dried blood spot (DBS) samples have been used for diagnostic purposes since their introduction in the neonatal screening of phenylketonuria almost 50 years ago. The range of its application has been extended to modern approaches, such as next-generation sequencing (NGS) for molecular genetic testing. This study aimed to evaluate the use of a standardized organic method for DNA extraction from DBS samples in the diagnostic setting.Methods: The clinical applicability of the method was tested using 3 samples collected from a newborn screening project for lysosomal storage diseases, allowing the determination of the genotype of the individuals. DNA was extracted from 3 3-mm diameter DBS punches. Quality, purity, and concentration were determined, and method performance was assessed by standard polymerase chain reaction, restriction length polymorphism, Sanger sequencing, and targeted NGS.Results: Results were compared with the ones obtained from DNA samples extracted following the internally validated in-house extraction protocol that used 6 3-mm punches of DBS and samples extracted from whole blood.Conclusion: This organic method proved to be effective in obtaining high-quality DNA from DBS, being compatible with several downstream molecular applications, in addition to having a lower cost per sample

Alpha-Galactosidase A Levels in Colombian Males with End-Stage Renal Disease: Ten Years of Selective Screening in Dried Blood Spots

Abstract Fabry disease is a metabolic alteration linked to an enzymatic deficiency of Alpha-Galactosidase A, this disorder compromises the sphingolipid metabolism, leading to an accumulation of lysosomal globotriaosylceramide and is inherited in an X-linked recessive way. The diagnostic of this disease, in general, requires the confirmation of below-normal levels of Alpha-Galactosidase A obtained from dried blood spot (DBS) samples, followed by an assessment of the enzyme in leukocytes. We aimed to report the Alpha-Galactosidase A values obtained in Colombian males with end-stage renal disease (ESRD) screened using dried blood spot samples during ten years. This screening was performed with samples sent to the analysis center from 6156 patients between 2006- 2016. All patients with low levels in enzyme activity (compared to the control population) were sent to confirmation through enzyme analysis in isolated leukocytes. 26 males (0.42%) with low levels of Alpha-Galactosidase A were identified (Range 0.0 - 1.14 nmol/ml/hour, cut-off: 1.15), 22 patients were subsequently measured in isolated leukocytes having a confirmation of Fabry disease in 5 patients (0.08% of total male population) (Range: 0.3 -4.7 nmol/mg prot/h). These results are similar to those reported in studies with comparable characteristics being this the first reporting frequency of Fabry disease among Colombian males with end-stage renal disease.

Estudo de estabilidade de anfetaminas e produtos de biotransformação de cocaína e tetraidrocanabinol em amostras de urina seca em papel (dried urine spots) / Stability study of amphetamines, biotransformation products of cocaine and tetrahydrocannabinol in dried urine spots

O número de pessoas utilizando substâncias ilícitas de forma recreativa aumenta a cada ano, chamando a atenção de estudiosos de diversas áreas do conhecimento. Com isso, a demanda de exames toxicológicos exigida para trabalhadores, vítimas de crimes e esportistas também tem crescido. A amostra biológica mais utilizada para análises toxicológicas continua sendo a urina, visto que sua obtenção é menos invasiva, possibilita coletar grande volume de amostra e pode-se detectar substâncias até dias após ter ocorrido a exposição ou consumo. Entretanto, estas amostras necessitam de um grande volume físico para serem armazenadas e transportadas aos laboratórios, devendo ser mantidas em temperatura baixa e controlada para conservação. Outro ponto a se considerar é a quantidade de amostra insuficientemente coletada, ou extravasamento do conteúdo, contaminando outras amostras e muitas vezes, inviabilizando a análise. Uma alternativa recente para tais problemas é utilizar a técnica chamada de dried urine spots (DUS), onde poucos microlitros de urina são colocados em um papel absorvente e secos sob temperatura ambiente, preservando de agentes degradantes os componentes presentes na urina. Assim, o objetivo deste trabalho é avaliar a estabilidade das substâncias do presente estudo em alta temperatura, temperatura ambiente e em temperaturas de 4°C e -20°C. Para este fim, foi necessário desenvolver, validar e aplicar métodos de extração e determinação de anfetaminas e produtos de biotransformação de cocaína e tetraidrocanabinol carboxílico (THCCOOH) em amostras dried urine spot, utilizando cromatografia líquida acoplada à espectrometria de massas. Os picos foram identificados por UPLC-ESI-MS/MS, com tempo total de 5 mins utilizando fase A- água, formiato de amônio e 0,1% ácido fórmico, e B- metanol: acetonitrila (6:4) + 0,1% de ácido fórmico. A extração foi feita utilizando acetonitrila: metanol: acetona (1:1:1) +ácido fórmico 0,1%. Não foi possível iniciar a validação de THCCOOH, visto uma possível complexação do analito com o papel. Para as outras substâncias, o método cromatográfico desenvolvido se mostrou eficiente e seletivo, com LOD e LOQ de 10 ng/mL para todos os analitos, sendo linear até 1000 ng/mL, atendeu as especificações de precisão e exatidão e carryover. As amostras permaneceram estáveis ao longo de 32 dias nas temperaturas estudadas, demonstrando a segurança em se utilizar a técnica de DUS para armazenamento e transporte de amostras biológicas dentro da faixa de temperatura do estudo até 32 dias

The number of people using illegal substances in a recreational way increases each year, drawing the attention of scholars from different areas of knowledge. As a result, the demand for workplaces drug tests, toxicological tests for victims of crimes and dopping has also grown. The biological sample most used for toxicological tests remains urine, since obtaining it is less invasive, it is possible to collect a large volume of sample and it is possible to detect substances up to days after exposure or consumption has occurred. However, these samples require a large physical volume to be stored and transported to the laboratories, and must be kept at a low temperature for conservation. Another point to consider is the amount of sample insufficiently collected, or leakage of the content, causing contamination of other samples and often making the analysis unfeasible. A recent alternative to such problems is to use "dried urine spots" (DUS), where few microliters of urine are placed on absorbent paper and dried at room temperature, preserving the components present in the urine from degrading agents. Thus, the objective of this work is to evaluate the stability of the substances in this study at high temperature, room temperature and at temperatures of 4°C and -20°C. For this purpose, it was necessary to develop, validate and apply methods of extraction and determination of amphetamines and biotransformation products of cocaine and carboxylic tetrahydrocannabinol (THCCOOH) in dried urine spot samples, using liquid chromatography coupled to mass spectrometry (LC-MS). The peaks were identified liquid chromatography coupled to a mass spectrometer (UPLC-ESI-MS/MS), with a total time of 5 mins using phase A- water, ammonium formate and 0.1% formic acid, and B- methanol: acetonitrile (6:4) + 0.1% formic acid. Extraction was done using acetonitrile: methanol: acetone (1:1:1) + 0.1% formic acid. It was not possible to perform the validation of THCCOOH, given a possible complexation of the analyte with the paper. To the others substances, the chromatographic method developed proved to be efficient and selective, with LOD and LOQ of 10 ng/mL for all analytes, being linear up to 1000 ng/mL, meeting the specifications of precision and accuracy and carryover. The samples remained stable for 32 days at the temperatures studied, demonstrating the safety of using the DUS technique for storage and transport of biological samples until 32 days on temperature range studied

Genetic characteristics of thalassemia in newborns in Baisha Li Autonomous County, Hainan Province / 中华地方病学杂志

Objective:To understand the incidence and genetic characteristics of thalassemia in newborns in Baisha Li Autonomous County, Hainan Province, and to provide data support for government decision-making departments to formulate appropriate policies for prevention and control of thalassemia.Methods:With the help of Newborn Disease Screening Network of Hainan Province, samples of dry blood spots on the heels of newborns born in Baisha Li Autonomous County from January to June 2020 were collected based on the principle of informed consent. Fluorescent PCR melting curve method was used to detect the common types of thalassemia genes in Chinese population, and some samples were verified by the PCR + flow-through hybridization method. Samples of suspected new or rare mutations were sent to gene companies for sequencing analysis.Results:A total of 391 samples of neonatal dry blood spots were collected, and 252 samples with thalassemia genes were detected, the detection rate was 64.45% (252/391). Among them, 213 samples with α-thalassemia genes were detected, and the detection rate was 54.48% (213/391); 13 samples with β-thalassemia genes were detected, and the detection rate was 3.32% (13/391); 26 samples with α- and β-thalassemia genes were detected, and the detection rate was 6.65% (26/391). Among the above mentioned thalassemia genotypes, 1 case of rare type α-thalassemia -α 4.2/HKαα and 1 case of rare type β-thalassemia β CD39/β N were detected. According to ethnicity, 176 samples with thalassemia genes were detected in 238 Li samples, with a detection rate of 73.95% (176/238); 67 samples with thalassemia genes were detected in 137 Han samples, with a detection rate of 48.91% (67/137); 9 samples with thalassemia genes were detected in 16 other ethnic samples, with a detection rate of 56.25% (9/16). Conclusions:The detection rate of neonatal thalassemia genes is relatively high in Baisha Li Autonomous County, Hainan Province, and α-thalassemia is the most common. It is recommended that relevant government departments of Hainan Province should carry out genetic testing of neonatal thalassemia in Baisha Li Autonomous County as soon as possible to ensure the quality of life of the newborns.

Research Progress on the Application of Artificial Intelligence in Rehabilitation Medicine in China / 中国医学科学院学报

The development of science and technology and the increasing demand of rehabilitation have driven the integration between artificial intelligence and rehabilitation medicine.In this study,statistical methods,document visualization tools,and other analysis methods were used in the Citespace software to analyze China's research status of artificial intelligence in the field of rehabilitation medicine with the key words of co-occurrence,emergence,and clustering.The relevant research hot spots were then classified and expounded.The results demonstrated that the current hot spots of artificial intelligence related to rehabilitation medicine included robots,brain-computer interfaces,human-computer interaction,and motor imagery.According to the clustering of key words and literature analysis,the five themes of artificial intelligence in rehabilitation medicine were determined as robot,brain-computer interface,intelligent rehabilitation training system,human-computer interaction,and assisted diagnosis and remote rehabilitation.Robotics and human-computer interaction would still be the research hot spots in the long future,and brain-computer interfaces,motor imagery,and remote rehabilitation would be new ones.This study analyzed the current hot spots,predicted the development trends,discussed the limitations,and proposed suggestions,aiming to provide reference for other scholars focusing on the application of artificial intelligence in rehabilitation medicine.

How Long are Residual Newborn Screening Specimens Useful for Retesting when Stored in Suboptimal and Uncontrolled Conditions of Temperature and Humidity?

Abstract Residual DBS specimens from newborns diagnosed with Phenylketonuria, Congenital Hypothyroidism, Cystic Fibrosis, Congenital Adrenal Hyperplasia and Galactosemia collected within 1995-2018, stored in cardboard boxes at ambient temperature in uncontrolled conditions, were retested for phenylalanine (Phe), thyrotropin (TSH), immunoreactive trypsinogen (IRT), total galactose (TGal) and 17-hydroxyprogesterone (17OHP), to demonstrate how long are they stable in these conditions and useful to reconfirm a previous abnormal result. Recovery percentage at retesting and qualitative interpretation regarding the current cutoff were evaluated. Phe, TSH and IRT recoveries showed decreasing trends along time. Phe recovery was 64 % after 2-years storage; TSH decayed rapidly recovering 47.3 % at 1-year, while IRT showed recoveries of 60 % at 1-year. Although 17OHP recovery presented a wide variation of results, a decaying trend was also found. Results suggest 17OHP is more stable than TSH and IRT, as supported by recoveries > 71 % when stored ≤ 2-years. TGal recovery presented an erratic behavior, so that it was not possible to estimate expected concentrations as a function of storage time. TGal recoveries above 100 % were found in UDP-galactose-4-epimerase and galactose-1-phosphate uridyltransferase deficiencies, evidencing possible galactose liberation from other sources. These results make a very valuable contribution for programs storing residual DBS in uncontrolled conditions.

Detecção de esteroides anabólicos androgênicos e seus produtos de biotransformação em dried urine spots por ionização por paper spray acoplada à espectrometria de massas (PS-MS) / Detection of anabolic androgenic steroids and its biotransformation products in dried urine spots by paper spray-mass spectrometry ionization (PS-MS)

Os esteroides anabólicos androgênicos (EAA) são utilizados clinicamente para tratar diferentes doenças, porém propagou-se o uso não terapêutico por atletas de elite e fisiculturistas, com o intuito de aumentar a massa muscular e melhorar o desempenho físico. O uso de substâncias ergogênicas, como fármacos estimulantes e narcóticos analgésicos no esporte, foi proibido em 1967 pelo International Olympic Committee (COI), mas somente em 1976 os EAA entraram para a lista de substâncias proibidas. O uso de EAA está associado a diversos efeitos adversos, principalmente cardiovasculares, neuroendócrinos e distúrbios psiquiátricos, além de dislipidemia, elevação dos marcadores inflamatórios e disfunção endotelial. As análises toxicológicas constam como a maneira mais eficaz de minimizar o doping no esporte. O material é fornecido pelo atleta durante as competições ou treinamentos e previne que os competidores alcancem vantagem competitiva devido ao uso de EAA. A utilização de métodos para amostragem alternativos tem ganhado força, devido à necessidade de técnicas mais práticas que utilizam pouco volume de amostra e possuem facilidade de armazenamento. O dried urine spots é um método no qual pequenas amostras de urina são aplicadas em papéis de filtro para análises qualitativas ou quantitativas. Ele se caracteriza por ser uma técnica rápida, fácil, simples e barata para a coleta, armazenamento e distribuição, além de minimizar os riscos de infecção, podendo ser utilizado na rotina. A técnica de paper spray (PS-MS) foi desenvolvida a partir da relevância de métodos como o dried blood spots por proporcionar análises mais rápidas, apresenta alta especificidade, capacidade de analisar diferentes analitos simultaneamente, baixo limite de detecção e dispensa a necessidade de reagentes específicos. Sendo assim, neste trabalho foi desenvolvido e validado o método de screening de EAAs em dried urine spots por ionização por paper spray acoplada à espectrometria de massas. O método apresentou limites de detecção entre 2-15ng/mL e presença de três interferentes endógenos. Os dez analitos de interesse deste estudo são estáveis por 150 dias em temperatura ambiente. Dessa forma, a análise de EAAs em dried urine spots por PS-MS demonstra grande potencial para se tornar um método alternativo no monitoramento rápido de drogas de abuso

Anabolic androgenic steroids (AAS) are used clinically to treat different diseases, but non-therapeutic use has spread among elite athletes and bodybuilders, with the aim of increase muscle mass and improve physical performance. The International Olympic Committee (IOC) banned the use of ergogenic substances, such as stimulating drugs and analgesic narcotics in sports, in 1967, but only in 1976, AAS were included on the list of prohibited substances. The use of AAS is associated with several adverse effects, mainly cardiovascular, neuroendocrine and psychiatric disorders, in addition to dyslipidemia, elevated inflammatory markers and endothelial dysfunction. Toxicological analyzes are the most effective approach to minimize doping in sport. The material is provided by the athlete during competitions or training and prevents competitors from achieving a competitive advantage due to the use of AAs. The use of alternative sampling methods has gained strength, due to the need for more practical techniques that use low sample volume and can be easily storage. Dried urine spots are a method, which a small amount of urine samples is applied to filter papers for qualitative or quantitative analysis. It is characterized by being a fast, easy, simple and inexpensive technique for collection, storage and distribution, in addition to minimizing the risks of infection, and can be used in the routine. The paper spray technique (PS-MS) was developed based on the relevance of methods such as dried blood spots for providing faster analysis, high specificity, ability to analyze different analytes simultaneously, low detection limit and for eliminating the need for specific reagents. Therefore, this work developed and validated a screening method for AAS in dried urine spots by paper spray-mass spectrometry ionization. The method provided detection limits between 2-15ng/mL and the presence of three endogenous interferents. The ten analytes of interest in this study are stable for 150 days at room temperature. Thus, the analysis of AAS in dried urine spots by PS-MS demonstrates great potential to become an alternative method for the rapid monitoring of drugs of abuse

Hypothermia Deaths in Van Province, Turkey

ABSTRACT Objective: To evaluate autopsy findings in hypothermia-related deaths in Van Province, Turkey, a city near the Turkey-Iran border. Methods: Autopsy reports on 43 hypothermia fatalities were retrospectively reviewed. Data regarding age, gender, nationality of the cases, seasonality of the deaths, crime scene findings, autopsy findings, manner of deaths, risk factors for hypothermia, other traumatic lesions, and toxicology were obtained from autopsy records and scene investigation records. Results: There were 36 males and 7 females. The mean age was 20.5 years. More than half of the cases died or were found dead in the spring months. All but one of the cases was found dead outdoors. There were common red-coloured livor mortis in 33 cases (76.7%), antemortem traumas in 16 cases (37.2%), cold erythema in 26 cases (60.5%), myxedema in 1 case, bloody discolouration in the synovial fluid in 11 (84.6%) cases, and Wischnewski spots in 32 cases (74.4%). Conclusion: The study showed that illegal refugees are an important social problem in Turkey. Hypothermia should be considered as a cause of death for refugees when they are found, especially in the cold provinces. In the diagnosis of hypothermia, bloody discolouration of the synovial fluid is confirmed to be a valuable finding. Wischnewski spots remain valuable for positive identification.

Fundamental pathological mechanisms underlying gastro-intestinal cancer peritoneal metastasis / 中华胃肠外科杂志

Gastrointestinal cancer peritoneal metastasis(GICPM) is one of the biggest challenges of clinical treatment. The ultimate solution to the problem requires the clinicians to accurately understand cytologic and molecular pathological mechanisms behind GICPM, and apply such knowledge in the clinical decision-making process for diagnosis and treatment of individual patient, so as to realize "prevention" and "treatment" proactively. The core cytopathological mechanisms behind GICPM, which are closely related to clinical treatment decisions, are as follows: (1) free cancer cells or clusters in peritoneal cavity colonize the peritoneum, resulting in irreversible pathological damage to peritoneal mesothelial cells; (2) the colonized cancer cells further invade the specific structure of the peritoneal milky spots and initiate an accelerated invasive growth process; (3) the process of peritoneal interstitial fibrosis aggravates the structural destruction of the peritoneum; (4) the interaction between cancer cells and immune cells in the milk spots forms a permissive immune microenvironment that promotes the growth of peritoneal metastatic cancer. These four core cytopathological mechanisms are mutually causal and promote each other, forming a vicious circle of GICPM development. As long as clinicians accurately understand these four points, it is possible to grasp the opportunity of clinical diagnosis and treatment, change reactive and passive treatment into preventive and proactive treatment, and improve the clinical diagnosis and treatment landscape of GICPM.