Aging parameters of the accelerated aging procedure through D-Galactose induction

Background and Objectives@#Intraperitoneal injection (i.p.) of D-galactose (D-gal) accelerates aging and develops aging models. A low dose of long-term use and a high dose of short-term use of D-gal can induce natural aging in mice, like brain, cardiac, liver, renal, and skin aging, and erectile dysfunction. Our research aims to determine whether a high dose of short-term use of D-gal. i.p. in rats can induce natural aging and affect the following parameters: body weight (BW), Superoxide Dismutase (SOD), Vascular endothelial growth factor (VEGF), C-reactive protein (CRP), and myostatin.@*Methods@#A daily D-gal i.p. dose of 300 mg/ml/kg for seven days was carried out to induce aging parameters in the rats. After seven days, the body and gastrocnemius circumference of the rats were weighed, and biochemical analysis for SOD, VEGF, CRP, and myostatin in the blood plasma was done.@*Results@#The data obtained were analyzed using nonparametric statistics Friedman test and Mann-Whitney test. After the seven day-intervention, both the control (NaCl 0.9% i.p.) and the high dose of short-term use of D-gal i.p. groups showed no significant difference in the body weight and gastrocnemius circumference. However, D-gal administration could increase the blood plasma level of SOD, VEGF, CRP, and myostatin.@*Conclusion@#We conclude that a high dose of short-term intraperitoneal D-galactose can be administrated to induce aging in rat models. The SOD, VEGF, CRP and myostatin can be used as aging parameters.

Validación de la reacción de polimerasa en cadena en tiempo real como herramienta diagnóstica en la aspergilosis asociada a COVID-19 en pacientes hospitalizados en Clínica Dávila de Santiago, Chile, durante los años 2020 y 2021 / Validation of real-time PCR as a diagnostic tool in aspergillosis associated with COVID-19 in patients hospitalized at Clínica Dávila at Santiago, Chile, during the years 2020 and 2021

INTRODUCCIÓN: Desde el inicio de la pandemia por COVID-19 se han registrado casos de infecciones de aspergilosis pulmonar asociada a esta infección, la cual tiene características diferentes a la aspergilosis pulmonar clásica y, por lo tanto, han significado un desafío diagnóstico. OBJETIVO: Validar una reacción de polimerasa en cadena (RPC) en tiempo real (sigla en inglés RT-PCR) comercial, como herramienta diagnóstica alternativa a la técnica de galactomanano (GM) en el diagnóstico de aspergilosis pulmonar asociada a COVID-19 (sigla en inglés CAPA). PACIENTES Y MÉTODO: Se analizaron resultados de RT-PCR de Aspergillus spp y GM en lavado bronco-alveolar (LBA) de 72 pacientes, hospitalizados por COVID-19 de Clínica Dávila entre los años 2020 y 2021. De estos pacientes, 33 presentaron CAPA. RESULTADOS: La RT-PCR de Aspergillus y GM presentaron una correlación positiva (r = 0,6351, valor p < 0,0001). La técnica de RT-PCR presentó una sensibilidad (S), especificidad (E), valor predictor positivo (VPP) y valor predictor negativo (VPN) de 100, 44, 66 y 100%, respectivamente, mientras que en GM fueron de 64, 89, 84 y 73%, respectivamente para LBA. Al utilizar ambas técnicas en combinación se obtuvo una S, E, VPP y VPN de 100, 82, 88 y 100%, respectivamente. CONCLUSIÓN: Este estudio concluyó que usar una técnica de RT-PCR de Aspergillus y GM en conjunto en LBA mejoraron los parámetros de desempeño de ambas técnicas usadas de manera individual para diagnosticar CAPA. Se requieren más estudios para evaluar el desempeño de técnicas combinadas en otros tipos de aspergilosis.

BACKGROUND: Since the beginning of the COVID-19 pandemic, there have been cases of pulmonary aspergillosis infections associated with this infection, which has different characteristics from classical pulmonary aspergillosis and therefore, have been diagnostic challenges. AIM: To validate a commercial real-time PCR (RT-PCR) method as an alternative diagnostic tool to the galactomannan (GM) technique in the diagnosis of COVID-19-associated pulmonary aspergillosis (CAPA). METHODS: Results of RT-PCR of Aspergillus spp and GM in broncho-alveolar lavage (BAL) of 72 patients hospitalized for COVID-19 at Clínica Dávila between 2020 and 2021 were analyzed. Of these patients, 33 presented CAPA. RESULTS: The RT-PCR for Aspergillus and GM showed a positive correlation (r = 0.6351, p-value < 0.0001). The RT-PCR for Aspergillus technique presented a sensitivity (S), specificity (S), positive predictive value (PPV) and negative predictive value (NPV) of 100, 44, 66 and 100% respectively, while the GM technique presented 64, 89, 84 and 73%, respectively for BAL. Using both techniques in combination a S, E, PPV and NPV of 100, 82, 88 and 100% were obtained respectively. CONCLUSION: This study concluded that using RT-PCR and GM techniques in combination in BAL improved the performance parameters of both techniques from those used individually to diagnose CAPA. Further studies are required to evaluate the performance of combined techniques in other aspergillosis focus.

Study of Cerebellar Tissue Following Induction of Aging in Mice By Di-Galactose / Estudio del Tejido Cerebeloso tras la Inducción del Envejecimiento en Ratones por D-Galactosa

SUMMARY: The cerebellum is a crucial area of the hindbrain that plays an essential role in balancing, excitement control, and subtle and accurate functions. Studies have shown that long-term use of D-galactose in mice, as with the symptoms of aging, causes morphological and functional disorders in the brain. This study was performed to evaluate the changes in the cerebellum cortex tissue and the measurement of reactive oxygen species (ROS) in the cerebellum following the induction of aging in mice by D-galactose. Accordingly, subjects were randomly assigned into two groups: Normal saline group and Aging group (D-galactose). To create an aging model, D- galactose, and saline solution (sodium chloride 0.9 %) were used. After completing the preparation and passage of the tissue, the cerebellum specimens were cut in 5 microns thickness and then stained with hematoxylin-eosin stain and finally examined under a Nikon microscope. Quantitative variables were analyzed by SPSS software using T-test. In the observations of cerebellum tissue samples, in the aged induced group by D-galactose, the most changes were observed in the Neuron purkinjense (Purkinje cells) layer. In the observations of the cerebellum tissue samples of aging group induced by D-galactose, the most changes were observed in the Neuron purkinjense, and the arrangement and placement of these cells were disorientated. The nucleus positioning was not central, and the Neuron purkinjense induced by aging were seen in different morphological forms. Necrosis, Chromatolysis, and Pyknosis were found. Based on the results, D-galactose (induction of aging) causes pathological changes in the cerebellar cortex, especially in the Neuron purkinjense layer.

El cerebelo es un área crucial del rombencéfalo que desempeña un papel esencial en el equilibrio, el control de la excitación y las funciones sutiles y precisas. Los estudios han demostrado que el uso a largo plazo de D-galactosa en ratones, al igual que con los síntomas del envejecimiento, provoca trastornos morfológicos y funcionales en el cerebro. Este estudio se realizó para evaluar los cambios en el tejido de la corteza del cerebelo y la medición de especies reactivas de oxígeno (ROS) en el cerebelo luego de la inducción del envejecimiento en ratones por D-galactosa. En consecuencia, los sujetos fueron asignados aleatoriamente a dos grupos: grupo de solución salina normal y grupo de envejecimiento (D-galactosa). Para crear un modelo de envejecimiento, se utilizaron D-galactosa y solución salina (cloruro de sodio al 0,9 %). Después de completar la preparación y el paso del tejido, las muestras de cerebelo se cortaron en un grosor de 5 µm y luego se tiñeron con tinción de hematoxilina-eosina y finalmente se examinaron bajo un microscopio Nikon. Las variables cuantitativas se analizaron mediante el software SPSS utilizando la prueba T. En las observaciones de muestras de tejido de cerebelo, en el grupo envejecido inducido por D-galactosa, la mayoría de los cambios se observaron en la capa de neuronas purkinjenses (células de Purkinje). En las observaciones de las muestras de tejido del cerebelo del grupo de envejecimiento inducidas por D-galactosa, la mayoría de los cambios se observaron en las neuronas purkinjenses, y la disposición y ubicación de estas células estaban desorientadas. El posicionamiento del núcleo no era central y las neuronas purkinjenses inducidas por el envejecimiento se observaban en diferentes formas morfológicas. Se encontró necrosis, cromatólisis y picnosis. Según los resultados, la D-galactosa (inducción del envejecimiento) provoca cambios patológicos en la corteza cerebelosa, especialmente en la capa de neuronas purkinjenses.

Rational design of L-arabinose isomerase from Lactobacillus fermentum and its application in D-tagatose production / 生物工程学报

L-arabinose isomerase (L-AI) is the key enzyme that isomerizes D-galactose to D-tagatose. In this study, to improve the activity of L-arabinose isomerase on D-galactose and its conversion rate in biotransformation, an L-arabinose isomerase from Lactobacillus fermentum CGMCC2921 was recombinantly expressed and applied in biotransformation. Moreover, its substrate binding pocket was rationally designed to improve the affinity and catalytic activity on D-galactose. We show that the conversion of D-galactose by variant F279I was increased 1.4 times that of the wild-type enzyme. The Km and kcat values of the double mutant M185A/F279I obtained by superimposed mutation were 530.8 mmol/L and 19.9 s-1, respectively, and the catalytic efficiency was increased 8.2 times that of the wild type. When 400 g/L lactose was used as the substrate, the conversion rate of M185A/F279I reached a high level of 22.8%, which shows great application potential for the enzymatic production of tagatose from lactose.

Effect of Erjing Pills on alleviating neuroinflammation of AD rats based on TLR4/NF-κB/NLRP3 pathway and its mechanism / 中国中药杂志

This paper aimed to study the effect of Erjing Pills on the improvement of neuroinflammation of rats with Alzheimer's di-sease(AD) induced by the combination of D-galactose and Aβ_(25-35) and its mechanism. SD rats were randomly divided into a sham group, a model control group, a positive drug group(donepezil, 1 mg·kg~(-1)), an Erjing Pills high-dose group(9.0 g·kg~(-1)), and an Erjing Pills low-dose group(4.5 g·kg~(-1)), with 14 rats each group. To establish the rat model of AD, Erjing Pills were intragastrically administrated to rats for 5 weeks after 2 weeks of D-galactose injection. D-galactose was intraperitoneally injected into rats for 3 weeks, and then Aβ_(25-35) was injected into the bilateral hippocampus. The new object recognition test was used to evaluate the learning and memory ability of rats after 4 weeks of intragastric administration. Tissues were acquired 24 h after the last administration. The immunofluorescence method was used to detect the activation of microglia in the brain tissue of rats. The positive expressions of Aβ_(1-42) and phosphory protein Tau~(404)(p-Tau~(404)) in the CA1 area of the hippocampus were detected by immunohistochemistry. The levels of inflammatory factors interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and interleukin-6(IL-6) in the brain tissue were determined by enzyme-linked immunosorbent assay(ELISA). Toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB)/nucleotide-binding oligomerization domain-like receptors 3(NLRP3) pathway-associated proteins in the brain tissue were determined by Western blot. The results showed that as compared with the sham group, the new object recognition index of rats in the model control group decreased significantly, the deposition of Aβ_(1-42) and p-Tau~(404) positive protein in the hippocampus increased significantly, and the levels of microglia activation increased significantly in the dentate gyrus. The levels of IL-1β, TNF-α, and IL-6 in the hippocampus of the model control group increased significantly, and the expression levels of TLR4, p-NF-κB p65/NF-κB p65, p-IκBα/IκBα, and NLRP3 proteins in the hippocampus increased significantly. Compared with the model control group, the Erjing Pill groups enhanced the new object recognition index of rats, decreased the deposition of Aβ_(1-42) and the expression of p-Tau~(404) positive protein in the hippocampus, inhibited the activation of microglia in the dentate gyrus, reduced the levels of inflammatory factors IL-1β, TNF-α, and IL-6 in the hippocampus, and down-regulated the expression levels of TLR4, p-NF-κB P65/NF-κB P65, p-IκBα/IκBα, and NLRP3 proteins in the hippocampus. In conclusion, Erjing Pills can improve the learning and memory ability of the rat model of AD presumably by improving the activation of microglia, reducing the expression levels of neuroinflammatory factors IL-1β, TNF-α, and IL-6, inhibiting the TLR4/NF-κB/NLRP3 neuroinflammation pathway, and decreasing hippocampal deposition of Aβ and expression of p-Tau, thereby restoring the hippocampal morphological structure.

Structural characterization of PCP-Ⅰ from Poria as vaccine adjuvant and its hydrolytic oligosaccharide / 中国中药杂志

Poria is an important medical herb in clinic. The authors isolated a polysaccharide(PCP-Ⅰ) from Poria in previous studies, which is composed of galactose, mannose, fucose and glucose. PCP-Ⅰ exhibited significant adjuvant effects on H1N1 influenza vaccine, hepatitis B surface antigen and anthrax protective antigen, and its adjuvant activity was stronger than aluminium adjuvant. However, little is known about the chemical structure of PCP-Ⅰ at present. In this study, weak acid hydrolysis was used to obtain the backbone oligosaccharide of PCP-Ⅰ. Then periodate oxidation, Smith degradation, methylation analysis, Fourier transform infrared spectroscopy(FT-IR), nuclear magnetic resonance(NMR) and gas chromatography-mass spectrometry(GC-MS) were performed to investigate the chemical structural features of PCP-Ⅰ and its hydrolytic oligosaccharide(PCP-Ⅰ-hy-1). These results suggested that the backbone of PCP-Ⅰ was composed of galactose with α anomeric carbon and β anomeric carbon. The linking residues of galactan are(1→),(l→6) and(1→2,6).

Yigong Powder regulates CXCL12/CXCR4 signaling to reduce glutamate release and prevent cognitive decline in mouse model of aging / 中国中药杂志

This study aims to explore the effect of preventive administration of Yigong Powder on the learning and memory abilities of the mouse model of aging induced by D-galactose and decipher the underlying mechanism, so as to provide a basis for the application of Yigong Powder in the prevention and treatment of cognitive decline. Forty KM mice were randomized into control, model, donepezil(1.5 mg·kg~(-1)), and high-dose(7.5 g·kg~(-1)) and low-dose(3.75 g·kg~(-1)) Yigong Powder groups. The mice in other groups except the control group were injected with D-galactose(200 g·kg~(-1)) at the back of the neck for the modeling of aging. At the same time, the mice were administrated with corresponding drugs by gavage for one month. Morris water maze was used to examine the learning and memory abilities of the mice. Hematoxylin-eosin staining was employed to observe the pathological and morphological changes of the hippocampus. The immunofluorescence assay was employed to detect the expression of ionized calcium-binding adapter molecule 1(IBA1), glial fibrillary acidic protein(GFAP), chemokine C-X-C-motif ligand 12(CXCL12), chemokine C-X-C-motif receptor 4(CXCR4) in the hippocampus and observe the positional relationship between IBA1, GFAP, and CXCR4. Western blot was employed to determine the protein levels of extracellular regulated kinase(ERK), p-ERK, and tumor necrosis factor receptor 1(TNFR1). Enzyme-linked immunosorbent assay was employed to measure the levels of glutamate and tumor necrosis factor(TNF-α) in the brain tissue and the level of TNF-α in the serum and spleen. Yigong Powder significantly shortened the escape latency, increased the times crossing platforms, and prolonged the cumulative time in quadrants of the aging mice. It alleviated the nerve cell disarrangement, increased intercellular space, and cell degeneration or death in the hippocampus and reduced the pathology score of the damaged nerve. Moreover, Yigong Powder reduced the positive area of IBA1 and GFAP, reduced the levels of TNF-α in the brain tissue, serum, and spleen, and decreased spleen index. Furthermore, Yigong Powder decreased the average fluorescence intensity of CXCL12 and CXCR4, reduced CXCR4-positive astrocytes and microglia, down-regulated the protein levels of p-ERK/ERK and TNFR1, and lowered the level of glutamate in the brain tissue. This study showed that the preventive administration of Yigong Powder can ameliorate the learning and memory decline of the D-galactose-induced aging mice by regulating the immune function of the spleen and the CXCL12/CXCR4 signaling in the brain to reduce glutamate release. However, the mechanism of Yigong San in preventing and treating dementia via regulating spleen and stomach function remains to be studied.

Different processed products of Polygonati Rhizoma treat Alzheimer's disease in rats: urine metabolomics based on UPLC-Q/TOF-MS / 中国中药杂志

The study investigated the effects of different processed products of Polygonati Rhizoma(black bean-processed Polygonati Rhizoma, BBPR; stewed Polygonati Rhizoma, SPR) on the urinary metabolites in a rat model of Alzheimer's disease(AD). Sixty SPF-grade male SD rats were randomized into a control group, a model group, a donepezil group, a BBPR group, and a SPR group, with twelve rats in each group. Other groups except the control group were administrated with D-galactose injection(100 mg·kg~(-1)) once a day for seven weeks. The control group was administrated with an equal volume of normal saline once a day for seven consecutive weeks. After three weeks of D-galactose injection, bilateral hippocampal Aβ_(25-35) injections were performed for modeling. The rats were administrated with corresponding drugs(10 mL·kg~(-1)) by gavage since week 2, and the rats in the model and control group with an equal volume of double distilled water once a day for 35 continuous days. The memory behaviour and pathological changes in the hippocampal tissue were observed. The untargeted metabolites in the urine were detected by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q/TOF-MS). Principal component analysis(PCA) and orthogonal partial least square-discriminant analysis(OPLS-DA) were employed to characterize and screen differential metabolites and potential biomarkers, for which the metabolic pathway enrichment analysis was conducted. The results indicated that BBPR and SPR increased the new object recognition index, shortened the escape latency, and increased the times of crossing the platform of AD rats in the Morris water maze test. The results of hematoxylin-eosin(HE) staining showed that the cells in the hippocampal tissue of the drug administration groups were closely arranged. Moreover, the drugs reduced the content of interleukin-6(IL-6, P<0.01) and tumor necrosis factor-α(TNF-α) in the hippocampal tissue, which were more obvious in the BBPR group(P<0.05). After screening, 15 potential biomarkers were identified, involving two metabolic pathways: dicoumarol pathway and piroxicam pathway. BBPR and SPR may alleviate AD by regulating the metabolism of dicoumarol and piroxicam.

Evaluación de la prueba Aspergillus Galactomannan Ag VircliaR Monotest como alternativa a kit Platelia ™ Aspergillus EIA / Evaluation of the Aspergillus Galactomannan ag VircliaR Monotest test as an alternative to Platelia™ Aspergillus EIA kit

INTRODUCCIÓN: La prueba Aspergillus galactomannan Ag Virclia® (GM-VClia) es una técnica de galactomanano monotest, auto-matizada, basada en inmunoensayo quimioluminiscente (CLIA). OBJETIVO: Evaluar el desempeño del test de GM-VClia en muestras de suero y lavado bronquioalveolar (LBA) procesadas previamente con el kit Platelia™ Aspergillus EIA (GM-Plat). MATERIALES Y MÉTODOS: Se estudiaron 56 muestras de suero y 40 de LBA, correspondientes a un total de 59 pacientes (algunos con determinación de galactomamano en ambas muestras) con enfermedades pulmonares, hematológicas, LES, Covid-19 y tumores, entre otros. Trece pacientes tuvieron aspergilosis invasora (1 probada y 12 probables). RESULTADOS: La correlación entre ambos métodos para suero y LBA fue r = 0,8861 p < 0,0001 y r = 0,6368 p < 0,001, respectivamente. Hubo una concordancia global de 67,7% (65/96), siendo de 85,7% (48/56) en sueros y 42,5,0% (14/49) en LBA. Al subir el punto de corte en LBA por GM-VClia la concordancia aumentó a 85,7%. CONCLUSIONES: Se observó una mayor correlación y concordancia en sueros que en LBA. El kit GM-VClia presentó una mayor sensibilidad y valor predictor negativo (VPN), que el kit GM-Plat. Las desventajas de GM-VClia, la constituyen la categoría "dudoso", que dificulta la interpretación y que, con los puntos de corte actuales en LBA, la correlación con GM-Plat es menor. Las ventajas son su mayor sensibilidad, facilidad de procesamiento y una mayor rapidez en los resultados.

BACKGROUND: The Aspergillus Galactomannan Ag Virclia® (GMVClia) test is a monotest and automated galactomannan technique based on chemiluminescent immunoassay (CLIA). AIM: To evaluate the performance of the GM-VClia test in serum and bronchioalveolar lavage (BAL) samples previously processed with the Platelia ™ Aspergillus EIA kit (GM-Plat). METHODS: 56 samples of serum 40 from BAL (some of them with galactomaman determination in both samples), from patients with pulmonary diseases, hematological diseases, SLE, Covid-19 and tumors, among others, were studied. Thirteen patients had invasive aspergillosis (1 proven and 12 probable). RESULTS: The correlation between both methods for serum and BAL was r = 0.8861 p < 0.0001 and r = 0.6368 p < 0.001, respectively. There was a global concordance of 67.7% (65/96), being 85.7% (48/56) in sera and 42.5.0% (14/49) in BAL. By raising the cut-off point in LBA by GM-VClia, the agreement increased to 85.7%. CONCLUSION: A greater correlation and concordance was observed in sera than in BAL. The GM-VClia kit had a higher sensitivity and NPV than the GM-Plat kit. The disadvantages of GM-VClia are the "doubtful" category, which makes interpretation difficult and that with the current cut-off points in LBA the correlation with GM-Plat is lower. The advantages are its greater sensitivity, ease of processing and faster results.

Ershiwuwei Shanhu Pills regulate Akt/mTOR/GSK-3β signaling pathway to alleviate Alzheimer's disease mice / 中国中药杂志

The present study investigated the mechanism of the Tibetan patent medicine Ershiwuwei Shanhu Pills(ESP) in alleviating Alzheimer's disease in mice via Akt/mTOR/GSK-3β signaling pathway. BALB/c mice were randomly assigned into a blank control group, a model group, low(200 mg·kg~(-1)), medium(400 mg·kg~(-1)) and high(800 mg·kg~(-1)) dose groups of ESP, and donepezil hydrochloride group. Except the blank control group, the other groups were given 20 mg·kg~(-1) aluminum chloride by gavage and 120 mg·kg~(-1) D-galactose by intraperitoneal injection for 56 days to establish Alzheimer's disease model. Morris water maze was used to detect the learning and memory ability of mice. The level of p-tau protein in mouse hippocampus and the levels of superoxide dismutase(SOD), malondialdehyde(MDA), catalase(CAT), and total antioxidant capacity(T-AOC) in hippocampus and serum were detected. Hematoxylin-eosin staining and Nissl staining were performed for the pathological observation of whole brain in mice. TdT-mediated dUTP nick-end labeling(TUNEL) staining was employed for the observation of apoptosis in mouse cortex. Western blot was adopted to detect the protein levels of p-mTOR, p-Akt, and GSK-3β in the hippocampus. Compared with the model group, the ESP groups showcased alleviated pathological damage of the whole brain, decreased TUNEL positive cells, reduced level of p-tau protein in hippocampus, and risen SOD, CAT, and T-AOC levels and declined MDA level in hippocampus and serum. Furthermore, the ESP groups had up-regulated protein levels of p-mTOR and p-Akt while down-regulated protein level of GSK-3β in hippocampus. Therefore, ESP can alleviate the learning and memory decline and oxidative damage in mice with Alzheimer's disease induced by D-galactose combined with aluminum chloride, which may be related to Akt/mTOR/GSK-3β signaling pathway.

Aumento en la positividad del biomarcador galactomanano durante la pandemia de COVID-19 en la Quinta Región: Valparaíso / Increase in the positivity of the galactomannan biomarker during the COVID-19 pandemic in the Fifth Region: Valparaíso

INTRODUCCIÓN: La actual pandemia provocada por SARS-CoV-2 ha provocado una alta carga en la salud pública y privada. Se han descrito casos y series de aspergilosis invasora asociada a pacientes con COVID-19 en ventilación mecánica. OBJETIVO: Describir el aumento en la positividad del biomarcador galactomanano (GM) durante la pandemia de COVID-19 en la Quinta Región: Valparaíso. MATERIALES Y MÉTODO: Estudio descriptivo, retrospectivo. Se revisó la cantidad y los resultados de GM, tanto de lavado bronco-alveolar (LBA) como en suero y los cultivos de LBA enviados al laboratorio de Micología de la Universidad de Valparaíso, desde enero y hasta septiembre del año 2020; luego se compararon con los exámenes recibidos en el mismo período del año 2019. RESULTADOS: Se observó un aumento significativo de los GM realizados en LBA, concentrándose principalmente entre los meses de julio y septiembre. El 29% de las muestras del año 2020 tenía el antecedente de ser de pacientes con COVID-19. Del total de muestras positivas durante el año de la pandemia, 5/12 fueron en pacientes con COVID-19. CONCLUSIONES: Hubo un aumento significativo de los GM realizados en LBA durante la pandemia, concentrándose principalmente entre los meses de julio-septiembre.

BACKGROUND: The current pandemic due to SARS-CoV-2 has caused a high burden on health. Cases and series of invasive aspergillosis associated with COVID-19 patients (CAPA) on mechanical ventilation have been described. AIM: To describe the increase in the positivity of the galactomannan (GM) biomarker during the COVID-19 pandemic in the Fifth Region: Valparaíso. METHOD: Retrospective descriptive study. The GM results in both broncho-alveolar lavage (BAL) and serum and the BAL cultures that were sent to the Mycology Laboratory of the University of Valparaíso from January to September 2020 were reviewed; then they were compared with the examinations of the same period of 2019. RESULTS: There was a significant increase in GMs carried out in LBA during the pandemic, concentrating mainly between the months of July-September. CONCLUSIONS: There was a significant increase in GM carried out in LBA during the pandemic, concentrating mainly between the months of July-September.

Mala absorción de glucosa y de galactosa y su asociación con el síndrome de Down / Malabsorption of glucose and galactose and its relation with Down sydrome

Introducción: La malabsorción de glucosa y de galactosa es una enfermedad genética autosómica recesiva debida a una mutación que afecta al cotransportador de sodio-glucosa. Objetivo: Describir una asociación infrecuente entre el síndrome de Down y la mala absorción de glucosa y de galactosa. Presentación del caso: Niño desnutrido de 3 ½ meses de edad con síndrome de Down, de padres consanguíneos. Presentó precozmente diarrea explosiva, vómitos con deshidratación. Se mejoró tras la eliminación de la alimentación oral y la perfusión hidroelectrolítica y empeoró con la utilización de las sales de rehidratación oral y las fórmulas lácteas, sin proteínas de leche de vaca. El estudio de las heces mostró un pH fecal /5, presencia de glucosa, ionograma de las 24 horas fue: sodio 0,5 mEq (1-10), potasio 2,6 mEq (8-22) y el cálculo realizado para distinguir entre diferentes causas de diarrea dio aumentado: 168 mOsm/kg (50-125). Ante este cuadro clínico se consideró el diagnóstico de malabsorción de glucosa y de galactosa sobre todo tras la mejora de la sintomatología bajo dieta exclusivamente azucarada con fructosa. Conclusiones: Es importante tener en cuenta la malabsorción de glucosa y de galactosa dentro de los diagnósticos diferenciales de las diarreas acuosas congénitas. El diagnóstico precoz y la dieta adecuada con fructosa evitan deshidratación y malnutrición. La particularidad de nuestro caso es la asociación de la malabsorción de glucosa y de galactosa con el síndrome de Down, que, según nuestro conocimiento, es la primera vez que se describe y podría aumentar la morbilidad(AU)

Introduction: Malabsorption of glucose and galactose is a genetic autosomic recesive disease caused by a mutation that affects the co-transportator of sodium-glucose. Objective: To describe an unfrequent relation between the Down syndrome and the malabsortion of glucose and galactose. Case presentation: Undernourished child, 3 and half months old with Down syndrome from parents by blood. He early presented explosive diarrhea and vomiting with dehydration. He improved his state after elimination of oral feeding and a hydroelectrolitic perfusion, and his state worsened with the use of oral rehydration salts and dairy formula, even without cow´s milk proteins. The study of feces showed a fecal pH /5, presence of glucose, and the ionogram after 24 hours showed: sodium 0,5mEg (1-10), potasium 2,6 mEg (8-22) and the calculation made to identify the different causes of diarrhea was increased: 168 mOsm/kg (50-125). Having this clinical records, it was considered a diagnostic of glucose and galactose malabsorption, mainly after the improvement of the symptoms under a diet exclusively sugared with fructose. Conclusions: It is important to take into account glucose and galactose malabsortion in the differential diagnosis of congenital watery diahrrea. The early diagnosis and an adequate diet with fructose avoid dehytration and malnutrition. The particularity of this case is the relation of glucose and galactose malabsorption with Down syndrome, that according to our knowledge is the first time it is described and it can increase the morbility(AU)

Expression and significance of follicular helper T cells and galactose-deficient IgA1 in children with Henoch-Schönlein purpura / 中国当代儿科杂志

OBJECTIVE@#To study the role of follicular helper T (Tfh) cells and galactose-deficient IgA1 (Gd-IgA1) in the pathogenesis of childhood Henoch-Schönlein purpura (HSP) and the correlation between them.@*METHODS@#A total of 36 children with newly-diagnosed HSP were enrolled. They were divided into two groups: HSP nephritis (HSPN) group with 11 children and non-HSPN group with 25 children according to the presence or absence of HSPN. Another 15 children who underwent physical examination at the outpatient service were enrolled as the healthy control group. Flow cytometry was used to measure the proportion of Tfh cells (CD4CXCR5ICOS) in peripheral blood. ELISA was used to measure the levels of interleukin-21 (IL-21) and interleukin-6 (IL-6) in peripheral blood and the serum levels of IgA1 and Gd-IgA1. A Pearson correlation analysis was used to investigate the correlation of serum Gd-IgA1 concentration with Tfh cells and related factors expression in the children with HSP.@*RESULTS@#Both the HSPN and non-HSPN groups had significantly higher proportion of Tfh cells and expression levels of IL-21 and IL-6 in peripheral blood than the healthy control group (P<0.05). The HSPN group had significant increases in the above indices compared with the non-HSPN group (P<0.05). Both the HSPN and non-HSPN groups had significantly higher serum levels of IgA1 and Gd-IgA1 than the healthy control group (P<0.05). The HSPN group had significantly higher serum levels of IgA1 and Gd-IgA1 than the non-HSPN group (P<0.05). In the children with HSP, serum Gd-IgA1 level was positively correlated with Tfh cells proportion and IL-21 and IL-6 levels (P<0.05).@*CONCLUSIONS@#Tfh cells and related cytokines and serum Gd-IgA1 are involved in the development of HSP/HSPN. Tfh cells may mediate the increased production of Gd-IgA1.

Modeling of lactose enzymatic hydrolysis using Monte Carlo method

Background: Mathematical modeling is useful in the analysis, prediction, and optimization of an enzymatic process. Unlike the conventional modeling methods, Monte Carlo method has special advantages in providing representations of the molecule's spatial distribution. However, thus far, Monte Carlo modeling of enzymatic system is namely based on unimolecular basis, not suitable for practical applications. In this research, Monte Carlo modeling is performed for enzymatic hydrolysis of lactose for the purpose of real-time applications. Results: The enzyme hydrolysis of lactose, which is conformed to Michaelis­Menten kinetics, is modeled using the Monte Carlo modeling method, and the simulation results prove that the model predicts the reaction kinetics very well. Conclusions: Monte Carlo modeling method can be used to model enzymatic reactions in a simple way for real-time applications.

Diagnostic value of the combinations of bronchoalveolar lavage fluid galactomannan test and serum galactomannan test in invasive pulmonary aspergillosis / 中华危重病急救医学

OBJECTIVE@#To evaluate the accuracy and diagnostic value of bronchoalveolar lavage fluid galactomannan test (BALF-GM) combined with serum GM test on invasive pulmonary aspergillosis (IPA).@*METHODS@#190 cases of BALF-GM and 4 787 cases of serum GM specimens suspected of fungal infection in patients admitted to Affiliated Hospital of Jining Medical University from January 2016 to June 2018 were enrolled and analyzed. All patients were classified into clinically confirmed IPA, clinically diagnosed IPA, suspected IPA and excluded IPA according to the classification standard of Expert consensus on diagnosis and treatment of pulmonary mycosis. The coincidence rate of BALF and serum GM test results with clinical diagnosis was analyzed. Receiver operating characteristic (ROC) curve was performed, and the diagnostic value of BALF and serum GM test alone or in combination for IPA was evaluated. Subgroup analysis was performed in patients with normal or abnormal immune function, and the sensitivity and specificity of BALF and serum GM test were compared separately or jointly.@*RESULTS@#The positive rate of BALF-GM was 46.8% (89/190), and 10.4% (497/4 787) on serum GM. Among them, 156 patients were both tested on BALF and serum GM. There were 44 cases with both positive in BALF and serum GM, the coincidence rate of clinical definite was 93.2% (41/44). There were 34 cases with positive BALF-GM and negative GM test in serum, and the coincidence rate of clinical definite was 64.7% (22/34). There were 56 cases positive in serum GM and negative in BALF-GM, and the coincidence rate of clinical definite was 48.2% (27/56). BALF and serum GM tests were both negative in 22 cases, and the coincidence rate of exclusion diagnosis was 90.9% (20/22). ROC curve analysis showed that the diagnostic value of BALF-GM test combined with serum GM test for IPA was better than that of BALF-GM test or serum GM test alone [area under ROC curve (AUC): 0.992 vs. 0.983, 0.976]. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were 95.3%, 87.0%, 93.2% and 90.9%, respectively. Subgroup analysis showed that among 89 patients with positive BALF-GM test, 85 cases (95.5%) had normal immune function and 4 cases (4.5%) had unknown condition. Among 497 patients with positive serum GM test, 12 cases (2.4%) had normal immune function, 372 cases (74.9%) had abnormal immune function and 113 cases (22.7%) were uncertain. It was shown by ROC curve analysis that the sensitivity of positive BALF-GM test in diagnosis of IPA in patients with normal immune function was higher than that of positive serum GM test (95.6% vs. 88.9%), while the sensitivity of positive serum GM test in patients with abnormal immune function was higher than that of positive BALF-GM test (91.8% vs. 89.9%).@*CONCLUSIONS@#The results of BALF and serum GM tests are in good agreement with clinical diagnosis, and the combined detection of BALF and serum GM is more valuable for IPA diagnosis than single detection, especially for patients with unknown immune function.

Value of galactose-deficient IgA1 in the early diagnosis of Henoch-Schönlein purpura nephritis in children / 中国当代儿科杂志

OBJECTIVE@#To explore the value of galactose-deficient IgA1 (Gd-IgA1) in the early diagnosis of Henoch-Schönlein purpura nephritis (HSPN) in children.@*METHODS@#A total of 67 hospitalized children who were definitely diagnosed with HSPN between January and April 2018 and 58 hospitalized children with Henoch-Schönlein purpura (HSP) were enrolled in the study. Twenty children undergoing routine physical examinations served as controls. The levels of serum and urine Gd-IgA1 were determined using ELISA. The receiver operating characteristic curve was used to analyze the value of serum Gd-IgA1 and urine Gd-IgA1/urine creatinine ratio in the diagnosis of HSPN.@*RESULTS@#The level of serum Gd-IgA1 and urine Gd-IgA1/urine creatinine ratio in children with HSP or HSPN were significantly higher than those in healthy control group (P<0.01), with a significantly greater increase observed in children with HSPN (P<0.01). Serum Gd-IgA1 ≥1 485.57 U/mL and/or urine Gd-IgA1/urine creatinine ratio ≥105.74 were of favorable value in the diagnosis of HSPN. During the six-month follow-up of the 49 children with HSP, the incidence of HSPN was 47% (23/49), which included a 100% incidence in children with serum Gd-IgA1 ≥1 485.57 U/mL and a 73% incidence in children with urine Gd-IgA1/urine creatinine ratio ≥105.74.@*CONCLUSIONS@#Serum and urine Gd-IgA1 is of favorable clinical value in the early diagnosis of HSPN.

Molecular mechanisms of mycelium of Cordyceps sinensis ameliorating renal tubular epithelial cells aging induced by D-galactose via inhibiting autophagy-related AMPK/ULK1 signaling activation / 中国中药杂志

To explore the effects and molecular mechanisms of mycelium of Cordyceps sinensis(MCs)improving renal tubular epithelial cells aging induced by D-galactose,the renal proximal tubular epithelial cells(NRK-52E cells)of rats in vitro were divided into the normal group(N),the D-gal model group(D),the low dose of MCs group(L-MCs),the medium dose of MCs group(M-MCs)and the high dose of MCs group(H-MCs),and treated by the different measures,respectively.More specifically,the NRK-52E cells in each group were separately treated by 1%fetal bovine serum(FBS)or D-galactose(D-gal,100 mmol·L~(-1))or D-gal(100 mmol·L~(-1))+MCs(20 mg·L~(-1))or D-gal(100 mmol·L~(-1))+MCs(40 mg·L~(-1))or D-gal(100 mmol·L~(-1))+MCs(80 mg·L~(-1)).After the intervention for24 h or 48 h,firstly,the effects of D-gal on the protein expression levels of klotho,P27 and P16,the staining of senescence-associatedβ-galactosidase(SA-β-gal)and the activation of adenosine monophosphate activated protein kinase(AMPK)/uncoordinated 51-like kinase 1(ULK1)signaling in the NRK-52E cells were detected,respectively.Secondly,the effects of MCs on the activation of the NRK-52E cells proliferation were investigated,respectively.Finally,the effects of MCs on the protein expression levels of klotho,P27,P16and microtubule-associated protein 1 light chain 3(LC3),the staining of SA-β-gal and the activation of AMPK/ULK1 signaling in the NRK-52E cells exposed to D-gal were examined severally.The results indicated that,for the NRK-52E cells,D-gal could cause aging,induce the protein over-expression levels of the phosphorylated AMPK(p-AMPK)and the phosphorylated ULK1(p-ULK1)and activate AMPK/ULK1 signaling pathway.The co-treatment of MCs at the medium and high doses and D-gal could significantly ameliorate the protein expression levels of klotho,P27,P16 and the staining of SA-β-gal,suggesting the anti-cell aging actions.In addition,the cotreatment of MCs at the medium and high doses and D-gal could obviously improve the protein expression levels of LC3,p-AMPK,and p-ULK1,inhibit the activation of AMPK/ULK1 signaling and increase autophagy.On the whole,for the renal tubular epithelial cells aging models induced by D-gal,MCs not only has the in vitro actions of anti-aging,but also intervenes aging process by inhibiting autophagy-related AMPK/ULK1 signaling activation,which may be the novel molecular mechanisms of MCs protecting against aging of the renal tubular epithelial cells.

Study of mycelial polysaccharide from Paraisaria dubia of Ophiocordyceps gracilis asexual / 中国中药杂志

In order to provide a foundation for the development and application of Ophiocordyceps gracilis and increase the new resources of cordyceps,an asexual Paraisaria dubia was isolated from an O. gracilis fruit body. After 10 days of liquid fermentation,white globular mycelium and clear transparent fermentation were produced. The mycelium was extracted by hot water and precipitated with ethanol to obtain intracellular crude polysaccharide. The protein was deproteinized to obtain deproteinized polysaccharide. The intracellular pure polysaccharide was purified by Sepharose 4 B column chromatography and were analyzed by UV,IR,1 H-NMR,and13 CNMR data,as well as GC and HPLC. The results showed that the intracellular polysaccharide of P. dubia was composed of glucose,galactose and mannose with a molar ratio of 25. 54 ∶2 ∶1. It was a β-configuration glycosylic bond,containing pyranoside. The initial connection of polysaccharide was β(1→2)(1→4)(1→6) connection. This experiment provides a theoretical basis for the development and application of P. dubia.

Evaluation of Acute and Chronic Effects of D-Galactose on Memory and Learning in Wistar Rats

OBJECTIVE: D-galactose has been commonly used in rodent models to induce accelerated effects of aging, including those on learning, memory, and muscular tone and coordination. This is normally seen on chronic administration of D-galactose. However, there is minimal suggestive evidence on the short-term effects of the same. The aim of the study was to study the acute and chronic effects of D-galactose on learning and memory in Wistar rats. METHODS: Twenty four male Wistar rats were randomly assigned to the control, standard (rivastigmine), oral D-galactose (200 mg/kg/day) and subcutaneous D-galactose (200 mg/kg/day) for a total duration of 8 weeks. Effects on learning and memory were assessed at 2 weeks, 4 weeks and 8 weeks by Morris water maze model and passive avoidance testing. RESULTS: Both oral and subcutaneous D-galactose showed positive effects on learning and memory on acute dosing, whereas this beneficial effect was lost during chronic dosing. CONCLUSION: Short-term administration of D-galactose showed positive effects, while long-term administration nullified these effects.

Effects of deoxygedunin on Alzheimer-like pathologic dysfunction induced by D-galactose combined with AlCl / 中国应用生理学杂志

OBJECTIVE@#To investigate the effects of Deoxygedunin on Aβ deposition, learning memory, and oxidative stress induced by D-galactose combined with AlCl in model rats with Alzheimer's disease and its possible mechanism.@*METHODS@#Male SD rats were randomly divided into three groups (=12):control group, model group (AD) and intervention group (AD+Deo). Morris water maze test was used to detect learning/memory and cognitive function in rats.Glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and malondialdehyde (MDA) contents in homogenate of hippocampus were detected by enzyme-linked immunosorbent assay (ELISA).Tau protein expression in rat cerebral cortex was detected by immunohistochemistry.Western blot was used to detect the expressions of extracellular signal regulated kinase 1(ERK1), protein kinase B (PKB) and tropomyosin-related kinase B (TrkB) on TrkB signaling pathway.@*RESULTS@#The results of water maze test showed that D-galactose combined with AlCl induced a significant increase in the escape latency compared with the control group (<0.05).Deoxygedunin could reverse the increase of the escape latency of the model group (<0.05).On the 7th day after removal of the platform, the model group showed an increase in escape latency compared with the control group and the intervention group (<0.01), and the number of crossing platforms was declined (<0.05); The results of immunohistochemistry and ELISA showed that the expressions of Aβ and tau protein in the model group were increased significantly compared with those of the control group (<0.01).The activities of SOD and GSH-Px were decreased significantly and the content of MDA was increased significantly.Compared with the model group, Deoxygedunin could reverse the increase of the expressions of Aβ and tau protein (<0.01), the decrease of SOD and GSH-Px activities (<0.05) and the increase of the MDA content (<0.05).Western blot results showed that Deoxygedunin treatment reversed the decreased phosphorylation levels of TrkB, AKT and ERK1 in hippocampus of the model group.@*CONCLUSIONS@#Supplement of Deoxygedunin can significantly reverse Aβ deposition, oxidative stress and cognitive deficits by activating the TrkB signal transduction pathway, which suggest that Deoxygedunin may serve as a promising therapeutic candidate for attenuating AD-like pathological dysfunction induced by D-galactose combined with AlCl.