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Introducción: La citometría de flujo permite la cuantificación de las subpoblaciones de linfocitos con una elevada sensibilidad, especificidad y objetividad. Estas ventajas solo se logran con un proceso laborioso de diseño individualizado y controlado para cada experimento. Objetivo: Diseñar un protocolo de un solo tubo policromático de citometría flujo para inmunofenotipo linfocitario periférico. Métodos: Se realizó un estudio experimental in vitro con muestras de sangre periférica obtenidas de tres voluntarios sanos, en el Centro Nacional de Genética Médica, en marzo de 2019. El tubo se compuso de seis marcadores de linaje para identificar linfocitos B, T, natural killer y natural killer T. Se desarrolló un protocolo de lisis de hematíes sin lavado. Se emplearon anticuerpos monoclonales conjugados con fluorocromos. El punto óptimo de concentración correspondió al mayor índice de tinción y conservación de los porcentajes de positividad de cada población. Se realizó la construcción progresiva del tubo y se propuso una estrategia lógica de secuencia de ventanas para el análisis de datos. Resultados: Los marcadores seleccionados permitieron realizar correctamente el inmunofenotipo linfocitario periférico. En los cinco puntos de titulación se observaron buenas discriminaciones entre las señales positivas y negativas, excepto para el anti-CD56 que presentó una tendencia decreciente del índice de tinción. El volumen total de conjugados requeridos para la determinación de los 6 antígenos fue de 3,75 μL por tubo. Conclusiones: Se obtuvo un tubo policromático que permite el inmunofenotipo periférico de forma rápida y precisa por seis antígenos linfocitarios simultáneamente, con el empleo de pequeños volúmenes de conjugado y sangre(AU)

Introduction: Flow cytometry allows quantification of lymphocyte subpopulations with high sensitivity, specificity and objectivity. These advantages are only achieved through the hardworking process of individualized and controlled design for each experiment. Objective: To design a flow cytometry protocol of a single polychromatic tube for peripheral lymphocyte immunophenotype. Methods: An experimental in vitro study was carried out, in March 2019, with peripheral blood samples obtained from three healthy volunteers, at the National Center for Medical Genetics. The tube was made up of six lineage markers for identifying natural B and T lymphocytes, natural killers and natural killer T cells. A protocol was developed for red blood cell lysis without washing. Fluorochrome-conjugated monoclonal antibodies were used. The optimal point of concentration corresponded to the highest staining index and preservation of the positivity percentages of each population. Progressive tube construction was performed and a logical window sequence strategy was proposed for data analysis. Results: The chosen markers allowed to carry out correct peripheral lymphocyte immunophenotype. Good discriminations between positive and negative signals were observed at the five titration points, except for anti-CD56, which presented a decreasing trend in the staining index. The total volume of conjugates required for determination of the six antigens was 3.75 μL per tube. Conclusions: A polychromatic tube was obtained that allows to carry out peripheral immunophenotype quickly and precisely by six lymphocyte antigens simultaneously, with the use of small volumes of conjugate and blood(AU)

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PURPOSE: The aim of this study was to determine whether different radiotherapy (RT) fractionation schemes induce disparate effects on lymphocyte and its subsets in breast cancer patients. METHODS: 60 female patients diagnosed with breast cancer were recruited in this study after receiving modified radical mastectomy and were randomly divided into two groups. One group received irradiation at a standard dose of 50 Gy in 25 fractions and the other at a dose of 40.3 Gy in 13 fractions. Both total lymphocyte count and its composition were recorded at three timepoints: right before the radiation treatment (T0), immediately after the last fraction of radiotherapy (T1) and 6 months after irradiation therapy ended (T2). RESULTS: Both groups experienced temporal lymphopenia after finishing local radiation (T1) (13F T0 vs. T1 1570.6 ± 243.9 vs. 940.6 ± 141.8, **p < 0.01; 25F T0 vs. T1 1620.5 ± 280.2 vs. 948.5 ± 274.6, **p < 0.01), while the lymphocyte count recovered at follow-up time (T2), and the cell count in the hypofractionation group (13F) was higher than the standard fraction group (25F) (13F vs. 25F 1725.6 ± 225.6 vs. 1657.5 ± 242.4, *p < 0.05). With respect to the composition of lymphocyte, we found T cell, B cell, and NK cell reacted differently to different radiotherapy protocols. CONCLUSIONS: Different RT protocols impose different impacts on immunity, leading us to further explore the optimal radiotherapy regimes to synergy with immunotherapy.

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In order to investigate the immune enhancement effects of Ophiopogon japonicus polysaccharide Ophiopogon japonicus (OJPS) on Newcastle disease (ND) live vaccine, chickens vaccinated against ND live vaccine was orally administered with the OJPS at high, medium and low concentrations respectively. In negative control group, chickens were given orally equal volume of physiological saline. On day 14, 21 and 28, the serum antibody titer, erythrocyte-C3b receptor rosette rate (E-C3bRR), erythrocyte-C3b immune complex rosette rate (E-ICRR) and peripheral lymphocyte proliferation were measured. The results showed that at most time points, the antibody titer, peripheral lymphocyte proliferation, E-C3bRR and elimination rate of immune complex of three OJPS administrating groups were significantly higher (P<0.05) than those in negative control group. It indicated that OJPS could significantly improve the immune efficacy of Newcastle disease live vaccine, Ophiopogon japonicus polysaccharide possessed synergistical immunoenhancement.(AU)

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In order to investigate the immune enhancement effects of Ophiopogon japonicus polysaccharide Ophiopogon japonicus (OJPS) on Newcastle disease (ND) live vaccine, chickens vaccinated against ND live vaccine was orally administered with the OJPS at high, medium and low concentrations respectively. In negative control group, chickens were given orally equal volume of physiological saline. On day 14, 21 and 28, the serum antibody titer, erythrocyte-C3b receptor rosette rate (E-C3bRR), erythrocyte-C3b immune complex rosette rate (E-ICRR) and peripheral lymphocyte proliferation were measured. The results showed that at most time points, the antibody titer, peripheral lymphocyte proliferation, E-C3bRR and elimination rate of immune complex of three OJPS administrating groups were significantly higher (P < 0.05) than those in negative control group. It indicated that OJPS could significantly improve the immune efficacy of Newcastle disease live vaccine, Ophiopogon japonicus polysaccharide possessed synergistical immunoenhancement.(AU)

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BACKGROUND: Neonatal lesion in the ventral hippocampus (NLVH) is a validated animal model to study schizophrenia from a neurodevelopmental perspective. This animal model is also used to investigate how neonatal lesions may alter the genetic expression of dopaminergic receptors. The present study compares mRNA expression levels of dopamine receptors (drd2 and drd3) in lymphocytes and brain of NLVH animals at two different age stages: young and adult. METHODS: The NLVH procedure was performed on 20 male Wistar rats at postnatal days 5-7. The mRNA expression levels of drd2 and drd3 genes in lymphocytes, nucleus accumbens, hippocampus and prefrontal cortex were measured and analyzed at postnatal days 45 and 90. The results were compared and contrasted with respective sham groups. RESULTS: In lymphocytes, only in NLVH-adult group we observed drd2 mRNA expression, while drd2 mRNA expression was not observed in the NLVH-juvenile rats; on the other hand, the drd3 mRNA expression did not show significant statistical differences. In hippocampus no differences were observed between drd2 mRNA or drd3 mRNA expression when comparing juvenile/adult shams with NLVH groups. In the prefrontal area, a decrease in drd2 mRNA expression levels were observed in the NLVH-adult group (F(1,3) = 52.83, p = 0,005) in comparison to the sham-adult group. Finally, in the nucleus accumbens, a strong decrease of drd3 mRNA expression was observed in the NLVH-adult group in comparison to the sham-adult group (F(1,3) = 123,2, p < 0.001). CONCLUSIONS: Our results show that differences in drd2 and drd3 mRNA levels in NLVH-adults are patent when compared to the sham-adult group or with the NLVH-juvenile group. These findings suggest that the expression levels may be regulated during adulthood, leading to behavioral and neurochemical changes related to schizophrenia. Therefore, more studies are necessary to determine the role of dopamine receptors as possible molecular markers for neurodevelopmental changes associated with schizophrenia.

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Background: Newcastle disease (ND), caused by avian paramyxovirus serotype 1 (APMV-1), also known as Newcastle disease virus (NDV), is regarded as one of the two most devastating diseases of poultry with the characteristics of serious disease and high flock mortality. It causes severe economic losses in domestic poultry, especially in chickens. At present, there were no effective drugs to treat this disease and the main method to control ND was vaccination. Moreover, new strains of virus resistant to chemotherapy continue to emerge, so does the need for a safe and effective vaccine. The immune adjuvant can make vaccine generate a strong immune response providing long-term protection against infection. With commonly usage of some immune adjuvants (e.g. mineral oil and aluminium hydroxide), many problems were occurred, such as side effects, strong local stimulation and carcinogenesis, together with complicated preparation, or failure to increase immunogenicity of weak antigen and so on. Botanical polysaccharides, as a new type of adjuvant or immunopotentiator, had become the hot research area because of their less side effects and no toxicity. The purpose of this research was to observe whether Chuanminshen violaceum polysaccharide (CVPS) possessed synergistical immunoenhancement, and offer the theoretical evidence for developing potential new-type adjuvant. Materials, Methods &a

Background: Newcastle disease (ND), caused by avian paramyxovirus serotype 1 (APMV-1), also known as Newcastle disease virus (NDV), is regarded as one of the two most devastating diseases of poultry with the characteristics of serious disease and high flock mortality. It causes severe economic losses in domestic poultry, especially in chickens. At present, there were no effective drugs to treat this disease and the main method to control ND was vaccination. Moreover, new strains of virus resistant to chemotherapy continue to emerge, so does the need for a safe and effective vaccine. The immune adjuvant can make vaccine generate a strong immune response providing long-term protection against infection. With commonly usage of some immune adjuvants (e.g. mineral oil and aluminium hydroxide), many problems were occurred, such as side effects, strong local stimulation and carcinogenesis, together with complicated preparation, or failure to increase immunogenicity of weak antigen and so on. Botanical polysaccharides, as a new type of adjuvant or immunopotentiator, had become the hot research area because of their less side effects and no toxicity. The purpose of this research was to observe whether Chuanminshen violaceum polysaccharide (CVPS) possessed synergistical immunoenhancement, and offer the theoretical evidence for developing potential new-type adjuvant. Materials, Methods &a

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Background: Newcastle disease (ND), caused by avian paramyxovirus serotype 1 (APMV-1), also known as Newcastle disease virus (NDV), is regarded as one of the two most devastating diseases of poultry with the characteristics of serious disease and high flock mortality. It causes severe economic losses in domestic poultry, especially in chickens. At present, there were no effective drugs to treat this disease and the main method to control ND was vaccination. Moreover, new strains of virus resistant to chemotherapy continue to emerge, so does the need for a safe and effective vaccine. The immune adjuvant can make vaccine generate a strong immune response providing long-term protection against infection. With commonly usage of some immune adjuvants (e.g. mineral oil and aluminium hydroxide), many problems were occurred, such as side effects, strong local stimulation and carcinogenesis, together with complicated preparation, or failure to increase immunogenicity of weak antigen and so on. Botanical polysaccharides, as a new type of adjuvant or immunopotentiator, had become the hot research area because of their less side effects and no toxicity. The purpose of this research was to observe whether Chuanminshen violaceum polysaccharide (CVPS) possessed synergistical immunoenhancement, and offer the theoretical evidence for developing potential new-type adjuvant. Materials, Methods & Results: 200 three-yellow chickens at one day of age were randomly divided into four groups and reared in separated pens. On 7 days old, the average maternal serum hemaglutination inhibition (HI) antibody titer was less than 3 log2, all chickens of each group (the average body weight (BW) was 120 g) were vaccinated with ND live vaccine through nasal drip and eye-drop. At the same time, the chickens in three CVPS groups (high, medium and low doses of CVPS) were orally administered with 0.5 mL of CVPS at concentrations of 100 mg/kg BW, 50 mg/kg BW, 25 mg/kg BW respectively, once a day for fi ve successive day, in negative control group (NC), with 0.5 mL of physiological saline, once a day for fi ve successive day. On days 14, 21 and 28, the serum antibody titer, erythrocyte-C3b receptor rosette rate (E-C3bRR), erythrocyte-C3b immune complex rosette rate (E-ICRR), peripheral lymphocyte proliferation and peripheral CD4+/CD8+ ratio were measured. The results showed that the antibody titer, E-C3bRR, elimination rate of immune complex and peripheral lymphocyte proliferation in three CVPS groups and peripheral CD4+/CD8+ ratio in medium dosage of CVPS group were significantly higher (P < 0.05) than those in control group throughout the process of whole experiment almost. Discussion: The CVPS not only improved the E-C3bRR and accelerated the elimination rate of CIC, but also induced higher antibody titer, peripheral lymphocyte proliferation and peripheral CD4+ /CD8+ ratio in chickens vaccinated against ND live vaccine. This indicated that CVPS possessed immune-enhancement efficacy of ND live vaccine and might be expected as a candidate of new-type adjuvant.

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Background: Newcastle disease (ND), caused by avian paramyxovirus serotype 1 (APMV-1), also known as Newcastle disease virus (NDV), is regarded as one of the two most devastating diseases of poultry with the characteristics of serious disease and high flock mortality. It causes severe economic losses in domestic poultry, especially in chickens. At present, there were no effective drugs to treat this disease and the main method to control ND was vaccination. Moreover, new strains of virus resistant to chemotherapy continue to emerge, so does the need for a safe and effective vaccine. The immune adjuvant can make vaccine generate a strong immune response providing long-term protection against infection. With commonly usage of some immune adjuvants (e.g. mineral oil and aluminium hydroxide), many problems were occurred, such as side effects, strong local stimulation and carcinogenesis, together with complicated preparation, or failure to increase immunogenicity of weak antigen and so on. Botanical polysaccharides, as a new type of adjuvant or immunopotentiator, had become the hot research area because of their less side effects and no toxicity. The purpose of this research was to observe whether Chuanminshen violaceum polysaccharide (CVPS) possessed synergistical immunoenhancement, and offer the theoretical evidence for developing potential new-type adjuvant. Materials, Methods &a

Background: Newcastle disease (ND), caused by avian paramyxovirus serotype 1 (APMV-1), also known as Newcastle disease virus (NDV), is regarded as one of the two most devastating diseases of poultry with the characteristics of serious disease and high flock mortality. It causes severe economic losses in domestic poultry, especially in chickens. At present, there were no effective drugs to treat this disease and the main method to control ND was vaccination. Moreover, new strains of virus resistant to chemotherapy continue to emerge, so does the need for a safe and effective vaccine. The immune adjuvant can make vaccine generate a strong immune response providing long-term protection against infection. With commonly usage of some immune adjuvants (e.g. mineral oil and aluminium hydroxide), many problems were occurred, such as side effects, strong local stimulation and carcinogenesis, together with complicated preparation, or failure to increase immunogenicity of weak antigen and so on. Botanical polysaccharides, as a new type of adjuvant or immunopotentiator, had become the hot research area because of their less side effects and no toxicity. The purpose of this research was to observe whether Chuanminshen violaceum polysaccharide (CVPS) possessed synergistical immunoenhancement, and offer the theoretical evidence for developing potential new-type adjuvant. Materials, Methods &a