S100 proteins and their implication in the thalamus: an integrative review / Las proteínas S100 y su implicación en el tálamo: una revisión integradora

SUMMARY: S100 proteins belong group of calcium-binding proteins and are present in physiological intracellular and extracellular regulatory activities, such as cell differentiation, and act in inflammatory and neoplastic pathological processes. Recently, its expressions in the nervous system have been extensively studied, seeking to elucidate its action at the level of the thalamus: A structure of the central nervous system that is part of important circuits, such as somatosensory, behavioral, memory and cognitive, as well as being responsible for the transmission and regulation of information to the cerebral cortex. This article is an integrative review of scientific literature, which analyzed 12 studies present in Pubmed. The analysis showed that the relationship of S100 proteins and the thalamus has been described in neoplastic processes, mental disorders, hypoxia, trauma, stress, infection, Parkinson's disease and epilepsy. In summary, it is possible to conclude that this protein family is relevant as a marker in processes of thalamic injury, requiring further studies to better understand its clinical, preclinical meanings and its prognostic value.

Las proteínas S100 pertenecen al grupo de proteínas fijadoras de calcio y están presentes en actividades reguladoras fisiológicas intracelulares y extracelulares, como la diferenciación celular, y actúan en procesos patológicos inflamatorios y neoplásicos. Recientemente, sus expresiones en el sistema nervioso han sido ampliamente estudiadas, buscando dilucidar su acción a nivel del tálamo: una estructura del sistema nervioso central que forma parte de importantes circuitos, como el somatosensorial, conductual, de memoria y cognitivo, así como además de ser responsable de la transmisión y regulación de la información a la corteza cerebral. Este artículo es una revisión integradora de la literatura científica, que analizó 12 estudios presentes en Pubmed. El análisis mostró que la relación de las proteínas S100 y el tálamo ha sido descrita en procesos neoplásicos, trastornos mentales, hipoxia, trauma, estrés, infección, enfermedad de Parkinson y epilepsia. En resumen, es posible concluir que esta familia de proteínas es relevante como marcador en procesos de lesión talámica, requiriendo más estudios para comprender mejor su significado clínico, preclínico y su valor pronóstico.

A case of neonatal-onset autoinflammation with infantile enterocolitis caused by a de novoNLRC4 gene variation / 中华围产医学杂志

This article reported a case of neonatal-onset autoinflammation with infantile enterocolitis (AIFEC) caused by NLRC4 gene mutation. The boy developed the disease in the neonatal period, presenting with recurrent fever, rash, hepatosplenomegaly and enterocolitis. Laboratory tests showed some indicators including ferritin and C-reactive protein were elevated. His condition was complicated by macrophage activation syndrome and anti-infective treatment was ineffective. High-throughput whole exome sequencing revealed a de novo heterozygous mutation of c.1021G>C (p.Val341Leu) in the NLRC4 gene and AIFEC was confirmed. AIFEC is a rare disease with no effective treatment at present, which can be developed in the neonatal period and diagnosed by whole exome sequencing.

Clinical significance of serum level changes of heart-type fatty acid-binding protein and S-100B protein in elderly patients with chronic heart failure / 中华老年医学杂志

Objective@#To investigate serum level changes of heart-type fatty acid-binding protein(H-FABP)and S100 calcium-binding protein B(S-100B)protein in elderly patients with chronic heart failure and their clinical significance.@*Methods@#A total of 160 patients with chronic heart failure treated at our hospital were recruited, and 80 healthy individuals receiving regular check-ups were enrolled as normal controls.Serum levels of H-FABP and S-100B and cardiac function index scores were compared between patients with different cardiac function grades.Correlations of serum H-FABP and S-100B levels with N-terminal pro-B-type natrlure tiepeptide(NT-proBNP)and with cardiac function index scores in heart failure patients were analyzed.The sensitivity, specificity and accuracy of serum H-FABP, S-100B and NT-proBNP for heart failure detection were compared.@*Results@#Serum levels of H-FABP, S-100B and NT-proBNP in elderly patients with chronic heart failure were elevated with increased cardiac function grading(F=9.823, 11.573 and 13.056, P=0.013, 0.000 and 0.000), and serum levels of H-FABP, S-100B and NT-proBNP were higher in elderly patients with heart failure than in the control group(P<0.05). Serum levels of H-FABP and S-100B were positively correlated with serum NT-proBNP levels, cardiac function grading and left ventricular end-diastolic diameter(LVEDd)(r=0.527, 0.510 and 0.487, P=0.008, 0.003 and 0.002; r=0.604, 0.496 and 0.533, P=0.006, 0.005 and 0.003), and were negatively correlated with left ventricular ejection fraction(LVEF)(r=-0.536 and-0.528, P=0.005 and 0.008). The sensitivity, specificity and accuracy of serum H-FABP combined with S-100B for heart failure detection were 93.2%, 91.6% and 95.7%, respectively.@*Conclusions@#Serum levels of H-FABP and S-100B are high in elderly patients with heart failure, and they are correlated with serum NT-proBNP levels, cardiac function grading and LVEDd.H-FABP combined with S-100B has a high positive rate for heart failure detection.

Clinical significance of serum level changes of heart-type fatty acid-binding protein and S-100B protein in elderly patients with chronic heart failure / 中华老年医学杂志

Objective To investigate serum level changes of heart-type fatty acid-binding protein(H-FABP)and S100 calcium-binding protein B(S-100B)protein in elderly patients with chronic heart failure and their clinical significance.Methods A total of 160 patients with chronic heart failure treated at our hospital were recruited,and 80 healthy individuals receiving regular check-ups were enrolled as normal controls.Serum levels of H-FABP and S-100B and cardiac function index scores were compared between patients with different cardiac function grades.Correlations of serum H-FABP and S-100B levels with N-terminal pro-B-type natrlure tiepeptide(NT-proBNP)and with cardiac function index scores in heart failure patients were analyzed.The sensitivity,specificity and accuracy of serum H-FABP,S-100B and NT-proBNP for heart failure detection were compared.Results Serum levels of H-FABP,S-100B and NT-proBNP in elderly patients with chronic heart failure were elevated with increased cardiac function grading (F =9.823,11.573 and 13.056,P =0.013,0.000 and 0.000),and serum levels of H-FABP,S-100B and NT-proBNP were higher in elderly patients with heart failure than in the control group(P＜0.05).Serum levels of H-FABP and S-100B were positively correlated with serum NT-proBNP levels,cardiac function grading and left ventricular end-diastolic diameter(LVEDd) (r =0.527,0.510 and 0.487,P =0.008,0.003 and 0.002;r =0.604,0.496 and 0.533,P =0.006,0.005 and 0.003),and were negatively correlated with left ventricular ejection fraction(LVEF) (r =-0.536 and-0.528,P =0.005 and 0.008).The sensitivity,specificity and accuracy of serum H-FABP combined with S-100B for heart failure detection were 93.2％,91.6％ and 95.7％,respectively.Conclusions Serum levels of H-FABP and S-100B are high in elderly patients with heart failure,and they are correlated with serum NT-proBNP levels,cardiac function grading and LVEDd.H-FABP combined with S-100B has a high positive rate for heart failure detection.

Expression of CCL5 and S100A4 protein in breast cancer tissues and their relationship with clinicopathological features and prognosis / 中国基层医药

Objective To investigate the relationship between the expression of CC chemokine ligand 5(CCL5)and S100 calcium binding protein A4 (S100A4)protein in breast cancer tissues with clinicopathological features and prognosis.Methods The expression of CCL5 and S100A4 in 40 cases of normal breast tissues and 120 cases of breast cancer were detected by immunohistochemistry,and the relationship between the degree of expres-sion and the clinicopathological features and prognosis of breast cancer was analyzed.Results The expression posi-tive rates of CCL5 and S100A4 in breast cancer tissues were 56.67% and 62.50% respectively,which were not expressed in normal breast tissues,and the differences were statistically significant (χ2CCL5 =39.403,P 0.05 ).And the expression of S100A4 was statistically correlated with clinical staging(χ2 =44.311,P 0.05).The expression of CCL5 and S100A4 in breast cancer was positively correlated(r =0.301, P <0.01).CCL5 was positively correlated with the recurrence of breast cancer(OR =6.270,P <0.01),and S100A4 was not correlated with the recurrence of breast cancer(OR =1.103,P =0.768).Survival analysis showed that the disease -free survival time of patients with positive CCL5 expression was significantly shorter than the patients with negative CCL5 expression(χ2 =11.851,P <0.01 ),and the disease -free survival time of patients with positive S100A4 expression was significantly shorter than the patients with negative S100A4 expression(χ2 =5.433,P =0.021).The joint detection showed that the disease -free survival time in CCL5(+)+S100A4(+)group was sig-nificantly lower than that of CCL5(+)or S100A4(+)group(χ2 =15.341,P <0.01)and CCL5 (-)+S100A4 (-)group(χ2 =15.341,P <0.01).Conclusion The expression of CCL5 and S100A4 in breast cancer can reflect the metastasis and staging of breast cancer,which can be used to judge the clinical pathological characteristics and prognosis of breast cancer.

La inmunorreactividad a calbindina en células de purkinje del cerebelo de ratones no es afectada por la infección con virus de la rabia / Immunoreactivity to calbindin in purkinje cells in the cerebellum of mice is not affected by rabies virus infection

Introducción: La calbindina (CB) es una proteína reguladora del calcio intracelular y la célula de Purkinje del cerebelo es la neurona con más alta concentración de CB. Se ha demostrado pérdida de inmunorreactividad a CB en diferentes áreas del sistema nervioso en ratones inoculados con virus de la rabia, pero faltaba estudiar este fenómeno en el cerebelo. Objetivo: Determinar el efecto de la inoculación con virus de la rabia sobre la expresión de CB en células de Purkinje del cerebelo de ratones. Metodología: Se inocularon ratones con el virus por vía intramuscular. Se sacrificaron los animales cuando alcanzaron la fase avanzada de la enfermedad y se fijaron mediante perfusión intracardiaca con paraformaldehído al 4%. Se les extrajo el cerebelo y se hicieron cortes sagitales de 50 micrómetros de espesor en un vibrátomo. Estos se procesaron mediante inmunohistoquímica para revelar la presencia de CB o de antígenos del virus de la rabia. El mismo procedimiento se realizó con animales no infectados (controles). Resultados: Las células de Purkinje fueron masivamente infectadas con el virus de la rabia. En las imágenes panorámicas observadas en el microscopio se comprobó que sólo estas células fueron inmunorreactivas a CB. No se hallaron diferencias significativas en la inmunorreactividad a CB, evaluada por densitometría óptica, entre los animales infectados y los controles. Conclusión: La expresión de CB en las células de Purkinje del cerebelo parece no afectarse por la infección con rabia, a diferencia de lo que se ha demostrado en otras áreas del sistema nervioso del ratón.

Introduction: Calbindin (CB) is a regulatory protein of intracellular calcium, and the cerebellar Purkinje cell is the neuron with the highest concentration of CB. Loss of CB immunoreactivity has been demonstrated in different areas of the nervous system in rabies virus-infected mice, but the study of this phenomena in the cerebellum lacked. Objective: To determine the effect of inoculation with rabies virus on the expression of CB in Purkinje cells of the cerebellum of mice. Methodology: Mice were intramuscularly inoculated with rabies virus. Animals were sacrificed when they reached an advanced stage of the disease and then they were fixed by intracardiac perfusion with 4% paraformaldehyde. Cerebellums were extracted and sagittal sections 50 microns thick were obtained in a vibratome. These were processed by immunohistochemistry to reveal the presence of CB protein or rabies virus antigens. The same procedure was performed with uninfected animals (controls). Results: Purkinje cells were massively infected with rabies virus. In the microscopic panoramic images observed was found that only these cells are immunoreactive to CB. No significant difference in CB immunoreactivity evaluated by optical densitometry was found between infected animals and controls. Conclusion: The expression of CB in Purkinje cells of the cerebellum appears not to be affected by infection with rabies unlike what has been shown in other areas of the mouse nervous system.

Expression of Jagged1 mRNA in human epithelial ovarian carcinoma tissues and effect of RNA interference of Jagged1 on growth of xenograft in nude mice / 中华妇产科杂志

Objective To investigate the expression of Jagged1 in human epithelial ovarian carcinoma tissues and the effect of Jagged1 on growth of xenograft in nude mice. Methods (1) Forty-eight cases of ovarian cancer and 30 cases of patients with benign epithelial ovarian tumor in the Henan Province Xinxiang Central Hospital during Feb. 2011 to Mar. 2014 were enrolled in this study. The mRNA expression of Jagged1, Notch1 and the downstream target genes Hes1, Hey1 were analyzed by using realtime PCR method. (2) The ovarian cancer xenograft models in nude mice were constructed by injecting SKOV3 cells in axillary subcutaneouswere. The nude mice were randomly divided into Jagged1 interference group, blank plasmid group and control group. Each group had 10 mice. They were transfected with pcDNA3.1(+)-siRNA-Jagged1, blank plasmid pDC3.1 and phosphate buffer, respectively. The tumor volumes and tumor masses were measured 14 days after transfection and the inhibition rate was calculated. The relative mRNA expression of Jagged1, Notch1, Hes1 and Hey1 in xenograft tissues after transfection in each group was detected by using realtime PCR technique and the relative protein expression of Jagged1, Notch1, Hes1 and Hey1 in xenograft tissues was detected by utilizing western blot method. Results (1) The relative mRNA expression of Jagged1, Notch1, Hes1 and Hey1 in ovarian cancer tissues were higher than benign ovarian tumor tissues, the differences were statistically significant (P0.05). Conclusions Jagged1 is highly expressed in epithelial ovarian carcinoma. Jagged1 gene interference in xenograft tumor can inhibit ovarian cancer cell growth and improve tumor suppressor rate, which probably play roles by inhibiting Notch1 signaling pathway.

Expression of S100A4 and hypoxia-induced factor 1α protein in type I endometrial carcinoma and their significances / 肿瘤研究与临床

Objective To investigate the expression of S100A4 and hypoxia-induced factor 1α (HIF-1α) protein in type Ⅰ endometrial carcinoma,and to analyze their relationship.Methods The expressions of S100A4 and HIF-1α protein were detected in 65 cases of type Ⅰ endometrial carcinoma and 40 normal endometrial tissues.Results The positive rate of S100A4 in endometrial carcinoma was 47.7 ％ (31/65),the normal tissues was 0 (0/40),and there was significant difference (x2 =27.069,P =0.001).The positive rate of HIF-lα was significantly higher in endometrial carcinoma [63.1％ (41/65)] than that in the normal tissues [7.5 ％ (3/40)] (x2 =31.417,P =0.001).The expressions of S100A4 and HIF-1α were positively related with FIGO staging,pathological grades,myometrial invasion and lymphatic metastasis (all P ＜ 0.05).The expressions of S100A4 and HIF-1α were positively correlated in type Ⅰ endometrial carcinoma (r =0.348,P =0.005).Conclusion The expression levels of S100A4 and HIF-1α are closely related with the oncogenesis and progression of type Ⅰ endometrial carcinoma,and their co-detection may be useful in prognosis assessment.

Human serum S100A12 level and severity of acute pancreatitis / 中华普通外科杂志

Objective To evaluate the correlation between human serum S100A12 level and severity of acute pancreatitis (AP).Methods Serum S100A12 were tested in 64 AP patients in 24 hours after the onset.S 100A12 levels was compared to the severity of AP,and the area under the curve (AUC) of the receiver operating characteristic curve(ROC) of serum S100A12 levels for estimating the severity of AP were made and compared with laboratory parameters and APACHE-Ⅱ,Ranson' s scoring system.Results S100A12 levels in early AP patients were higher than healthy controls (P ＜ 0.05).S100A12 levels increased and correlated with MCTSI scores:patients scored 7-8 ＞ patients scored 4-6 ＞patients scored less than 4 (P ＜ 0.01).S100A 12 levels increased with organ failure (P ＜ 0.01).The AUC of S100A12 in 24 hours after onset for distinguishing MAP between MSAP and SAP was 0.80,cut-off point was 61.83 ng/ml,sensitivity was 70.01％ and the specificity was 73.52％.Serum S100A12 levels were higher in SAP than in MSAP(P =0.01).The AUC of S100A12 in early AP for distinguishing between MSAP and SAP was 0.84,cut-off point was 285.32 ng/ml,the sensitivity was 76.94％ and the specificity was 94.12％,Youden index was 71.00％,positive likelihood ratio was 13.00,negative likelihood ratio was 0.20,the diagnostic performance was better than APACHE-Ⅱ,Ranson and Marshall scoring systems and serum CRP concentration.Conclusions Human serum S100A12 levels elevated at early stage of AP.S100A12 ＞285.32 ng/ml represents high risk of SAP,which is more sensitive and accurate than APACHE-Ⅱ and Marshall scoring system.

Human prefrontal layer II interneurons in areas 46, 10 and 24 / Interneuronas de la lámina II en las áreas prefrontales 46, 10 y 24 del humano

Background: Prefrontal cortex (PFC) represents the highest level of integration and control of psychic and behavioral states. Several dysfunctions such as autism, hyperactivity disorders, depression, and schizophrenia have been related with alterations in the prefrontal cortex (PFC). Among the cortical layers of the PFC, layer II shows a particular vertical pattern of organization, the highest cell density and the biggest non-pyramidal/pyramidal neuronal ratio. We currently characterized the layer II cytoarchitecture in human areas 10, 24, and 46. Objective: We focused particularly on the inhibitory neurons taking into account that these cells are involved in sustained firing (SF) after stimuli disappearance. Methods: Postmortem samples from five subjects who died by causes different to central nervous system diseases were studied. Immunohistochemistry for the neuronal markers, NeuN, parvalbumin (PV), calbindin (CB), and calretinin (CR) were used. NeuN targeted the total neuronal population while the rest of the markers specifically the interneurons. Results: Cell density and soma size were statically different between areas 10, 46, 24 when using NeuN. Layer II of area 46 showed the highest cell density. Regarding interneurons, PV+-cells of area 46 showed the highest density and size, in accordance to the proposal of a dual origin of the cerebral cortex. Interhemispheric asymmetries were not identified between homologue areas. Conclusion: First, our findings suggest that layer II of area 46 exhibits the most powerful inhibitory system compared to the other prefrontal areas analyzed. This feature is not only characteristic of the PFC but also supports a particular role of layer II of area 46 in SF. Additionally, known functional asymmetries between hemispheres might not be supported by morphological asymmetries.

Antecedentes: La corteza prefrontal (CPF) representa el nivel más alto de integración y control de funciones psíquicas y comportamentales. Varias patologías como autismo, desórdenes de hiperactividad, depresión y esquizofrenia se han relacionado con alteraciones de la CPF. La lámina II de las áreas que constituyen la CPF posee un patrón de organización vertical, una alta densidad celular y la mayor proporción de neuronas no-piramidal/piramidal. Sin embargo, la distribución del componente inhibitorio en estas regiones no se ha descrito. Objetivo: En el presente estudio nos propusimos caracterizar la lámina II de las áreas 10, 24 y 46 del humano, particularmente su componente inhibitorio teniendo en mente su participación en procesos de actividad sostenida relevantes cuando desaparece el estímulo. Métodos: Se utilizaron muestras de cinco sujetos que fallecieron por causas diferentes a enfermedades del sistema nervioso. Se tomaron secciones de las áreas 10, 24 y 46 de Brodmann y se procesaron con los anticuerpos contra NeuN para determinar la población neuronal total y contra Parvalbumina (PV), Calbindina (CB) y Calretinina (CR) para analizar la población de interneuronas. Resultados: Los resultados no mostraron diferencias interhemisféricas entre las áreas. Sin embargo, las tres áreas seleccionadas son significativamente diferentes entre sí en todos los parámetros analizados. El área 46 posee la mayor densidad y tamaño de interneuronas positivas para PV. Conclusiones: La ausencia de asimetrías morfológicas no permite explicar las asimetrías funcionales. La lámina II del área 46 posee el sistema inhibitorio más poderoso. Teniendo en cuenta la arquitectura modular de las capas supragranulares, este sistema inhibitorio subyace a la actividad sostenida, eje fundamental de la memoria operativa.

Effect of early application of naloxone on serum hs-CRP and S100B protein in neonates with hypoxic-ischemic encephalopathy / 中国基层医药

Objective To study the effect of early application of naloxone on serum high sensitivity C-reactive protein(hs-CRP) and S100B protein in neonates with hypoxic-ischemic encephalopathy.Methods According to the digital table,86 neonates with hypoxic-ischemic encephalopathy were randomly divided into the control group (n =43 cases) and the observation group (n =43 cases).The control group was treated by conventional therapy,while the observation group was treated by conventional therapy plus naloxone.They were treated for 7-10 days.The clinical effect was evaluated.Serum hs-CRP and S100B protein were detected before and after treatment.Results The total effective rate of the control group was 86.0％,which was significantly lower than 97.7％ of the observation group (x2 =5.78,P ＜ 0.05).The sense of recovery,reflecting the recovery and muscle tension recovery time of the observation group were less than those in the control group (t =5.017 2,3.991 6,2.904 8,all P ＜ 0.05).After treatment,the sertum hs-CRP and S100B protein in the two groups were significantly decreased (t =6.776 2,10.903 8,7.772 8,11.092 6,all P ＜ 0.01),compared with the control group,the decrease of the observation group was more significant (t =3.368 2,3.092 8,all P ＜ 0.05).Conclusion Early application of naloxone in neonates with hypoxic-ischemic encephalopathy can decrease serum hs-CRP and S100B protein.

Alterations in Calcium-Binding Properties of Sarcoplasmic Reticulum Membrane Proteins Following Cardiac Injury.

The objective of present work is to investigate metabolic alterations associated with heart failure, particularly one of its manifestations, a sustained hypocalcemia that causes hemodynamic changes contributed to subsequent myocardial injury. Comparative study was carried out using experimental models of pancreatic necrosis (PN) and crush syndrome (CS) accompanied by cardiac damage down to myocardial infarction. Study design: Wistar adult male rats randomly divided into groups (n=12/group). The controls are healthy intact animals. The pancreatic necrosis (PN) and crush syndrome (CS) groups were then randomly subdivided: PN group- into 3, 24 and 72 h groups concerning hemorrhage, early and late pancreatic necrosis respectively; CS group – into 2, 4, 24, and 48 h decompression stages. The rats were sacrificed to analyze spectra and calcium-binding properties of the membrane proteins isolated from the cardiomyocyte sarcoplasmic reticulum (SR). Development of pathological changes in the heart and pancreas were also monitored. Place and Duration of Study: Department of Pathological Biochemistry and Radioisotope Methods, H. Buniatyan Institute of Biochemistry of Natl. Acad. Sci (NAS), Republic of Armenia (RA). Experiments conducted between May 2011 and October 2013. Methodology: To study pathogenesis of hypocalcemia underlying myocardial damage a translocation of radioactive 45CaCI2 into cardiomyocytes and its intracellular distribution was examined. Binding of 45Ca2+ to the SR membrane proteins was measured after proteins separation by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE) and radioactivity from the gel plates was counted by a gas-flow meter Berthold–II. Isoelectric focusing of the protein isolated from the SR of cardiomyocyte was performed. Results: Statistically significant changes in mean radio labeled calcium incorporation into a total protein fraction of the cardiomyocyte SR from control (13682±271) were determined by3h of PN (23055±168, P<.001), 24 h of PN (22876±240, P<.01), and by 72 h (3851±271,P<.01), P vs. control. Similarly, these parameters were detected following CS by 2h decompression (24179±225, P<.01), 4-24 hours decompression (21666±124, P<.001) and 48 h decompression (2941±189, P<.001), P vs. control. We demonstrate that drop in the binding calcium level observed was partially due to impaired affinity to calcium of the cardiomyocyte SR calcium-binding proteins during development of both PN and CS despite a simultaneous manifestation of affinity to calcium of the SR 32-kDa protein. Conclusion: In the present study we have clearly shown that both experimental acute pancreatitis and long-term compression injury may cause similar changes,а loss the calcium-binding properties of the cardiomyocyte proteins, particularly those of SR serving as a main calcium depot under physiological circumstances and appear to be involved in common cellular and molecular mechanisms of myocardial injury contributing to hypocalcemia. Simultaneously, both PN and/or CS cause similar manifestations of the new calcium-binding properties of the cardiomyocyte SR 32-kDa membrane protein, and mirrored dynamic changes in its calcium affinity suggested by Scatchard plot analysis indicating a common mechanism that would be a transient attempt of certain heart cells to compensate hypocalcemia, and thus emerge from an otherwise pathological outcome. Thus, the above mentioned changes could be used to identify patients at high risk of cardiovascular disease in different pathologies.

Efecto de la degradación post mórtem sobre la detección inmunohistoquímica de antígenos en el cerebro de ratón / Effect of postmortem degradation on the immunohistochemical detection of antigens in the mouse brain

Introducción. Si bien lo ideal es llevar a cabo la preservación de los tejidos en el menor tiempo posible luego de la muerte de un animal objeto de un estudio neurohistoquímico, con frecuencia es inevitable trabajar con tejido nervioso obtenido varias horas post mórtem. Objetivo, Estudiar el efecto de la degradación post mórtem sobre la inmunorreacción de di-ferentes antígenos en el cerebro de ratón. Métodos. Se inocularon ratones con virus de la rabia y se extrajeron los cerebros luego de fijar los animales con paraformaldehído me-diante perfusión. En otro grupo de animales la extracción de los encéfalos se hizo para fi-jarlos por inmersión con el mismo fijador y en diferentes horas post mórtem. En un vibráto-mo se obtuvieron cortes coronales de los ce-rebros, y estos se procesaron para inmunode-tección de rabia y de otros cuatro antígenos. Resultados. Cuatro de los antígenos evalua-dos, calbindina, parvoalbúmina, glutamato y ácido gamma-aminobutírico (GABA), presen-taron pérdida de inmunorreacción cuando el tejido cerebral se había tratado previamente mediante fijación por inmersión. Este efecto fue más acentuado cuando aumentó el tiempo post mórtem antes de la fijación. Por el con-trario, la inmunorreacción al virus de la rabia se incrementó cuando transcurrieron más de seis horas post mórtem antes de la fijación. Conclusiones. La fijación por perfusión es ideal para estudios de inmunohistoquími-ca de diferentes antígenos. La degradación tisular post mórtem generalmente provoca disminución de la inmunorreacción. No obs-tante, los antígenos del virus de la rabia in-crementan su inmunorreacción a medida que transcurre el tiempo post mórtem antes de la fijación.

Introduction: It is advisable to carry out the preservation of tissues in the shortest time after the death of an animal subject of neu-rochemical study but it is often unavoidable to work with nervous tissue obtained several hours postmortem. Objective: To study the effect of postmor-tem degradation on immunoreactivity of di-fferent antigens in the mouse brain. Methods: Mice were inoculated with rabies virus and the brains were removed after the animals were fixed by perfusion with parafor-maldehyde. In another group of animals the brain extraction was performed and they were fixed by immersion in the same fixative solu-tion at different hours postmortem. Coronal sections of the brains were obtained in a vibra-tome and they were processed for immunode-tection of rabies, and other four antigens. Results: Four of the antigens studied, cal-bindin, parvalbumin, glutamate and GABA, showed loss of immunoreactivity when brain tissue was pretreated by immersion fixa-tion. This effect was more noticeable when postmortem time increase before the fixing. Conversely immunoreactivity to rabies virus was increased over six hours postmortem before fixation. Conclusions: Fixation by perfusion is ideal for immunohistochemical studies of diffe-rent antigens. Postmortem tissue degrada-tion usually causes decreased immunoreac-tivity. However, rabies virus antigens show increased immunoreactivity when elapses more postmortem time before fixation.

Excretion and clinical value of fecal calprotectin in preterm infants during the first two weeks of life / 国际儿科学杂志

Objective To determine the time course of fecal calprotectin (FC) excretion in preterm infants in the first two weeks of life and to identify influencing factors of FC levels.To explore whether FC may be an useful marker for the identification of gastrointestinal conditions in preterm infants.Methods FC concentration was measured using ELISA in 141 samples (50-100 mg)obtained from 47 preterm infants(gestational age less than 32 weeks or birth weight less than 1 200 grams)at birth,on day 7 and on day 14(The following was called FC1,FC2 and FC3 level for short).Results FC3 level was significantly higher than FC1 and FC2 levels (P ＜0.05),there were no significant differences between FC1 and FC2 levels in the 47 preterm infants(P ＞ 0.05).The FC1 level in infants whose mothers had antenatal infection was significantly higher(P ＜ 0.05).The FC2 level was negatively correlated with the antibiotic courses(r =-0.325,P ＜ 0.05).The FC2 level in infants with feeding intolerance was significantly lower than that in infants with feeding tolerance(P ＜ 0.05).The FC2 level in infants with feeding tolerance and the FC3 level in infants with feeding intolerance were positively correlated with the volume of enteral feeds,respectively (r =0.433,0.479,P ＜ 0.05).Contusion Antenatal infection and postnatal enteral feeds may lead to an increase in the excretion of FC in preterm infants.FC level was associated with enteral feeding and may be a useful marker for evaluation of gastrointestinal function in preterm infants.

Neuroprotective Effect of Sevoflurane on Controlled Hypotension in Patients with Coronary Heart Disease Undergoing Craniocerebral Surgery / 医药导报

Objective To study the neuroprotective effect of sevoflurane on controlled hypotension in patients with coronary heart disease undergoing craniocerebral surgery. Methods Twenty-six patients with coronary heart disease undergoing craniocerebral surgery were randomly divided into two groups,receiving either inhaled 2%-5%sevoflurane plus intravenous sodium nitroprusside (treatment group,n=13) or intravenous sodium nitroprusside 3-8 μg·kg-1 alone (contorl group,n=13) for blood pressure control. The hemodynamic changes were recorded during the operation. Patient satisfaction with surgeons and the duration hospital stay after surgery were recorded. The levels of cardiac troponin I (TNI),creatine kinase-MB (CK-MBM),neuron specific enolase ( NSE) and S100 calcium binding proteinβ( S-100βprotein) in serum were detected at one day pre-operation,the end of the operation,and one day post-operation. Results The duration of hospital stay after surgery was significantly shortened in treatment group [(20. 3±3. 8) versus (23. 9±4. 6) d,P<0. 05) compared with control group. The average heart rate significantly decreased, patient satisfaction significantly increased, and serum levels of NSE and S-100β protein one day post-operation significantly decreased as compared to control group (all P<0. 05). Compared with the day before operation,serum levels of NSE and S-100βprotein in the two groups increased significantly at the end of surgery (P<0. 05),and the levels of TNI,CK-MBM,NSE and S-100βsignificantly elevated one day post-operation (P<0. 05). Compared with the end of operation,serum levels of NSE and S-100βin contorl group incarcerated remarkably (P<0. 05) Conclusion Sevoflurane plays an important neuroprotective role,as evidenced by improving patients' satisfaction,reducing hospital stay after surgery,and maintaining the balance of myocardial oxygen delivery and consumption in craniocerebral surgery patients with coronary heart disease during controlled hypotension. However,it can not prevent postoperative myocardial injury in these patients.

Effects of flavonoids from Cuscuta chinensis on intestinal calcium-binding protein mRNA expression in ovariectomized osteoporosis model rats / 中国组织工程研究

BACKGROUND:Cuscuta chinensis is a mature seed of Cuscutachinensis Lam., can warm kidney. Previous studies demonstrated that kidney compound composed of Cuscuta chinensis could apparently inhibit bone loss and improve bone density. OBJECTIVE:To explore the influence of flavonoids from Cuscuta chinensis on bone mineral density of femur, 1,25(OH)2D3 levels of serum and kidney, the expression of smal intestine CaBp-D9K mRNA in model rats with ovariectomized osteoporosis. METHODS:A total of 72 Sprague-Dawley female rats were equal y and randomly divided into six groups (n=12):sham surgery group, model group, vitamin D3 group and low-, moderate-and high-dose flavonoids groups. The sham surgery group only received sham operation and the other five groups were ovariectomized respectively. One week after ovariectomy, the rats were given flavonoids from low-, moderate-and high-dose Cuscuta chinensis and vitamin D3 (2 mg/kg) by intragastric administration for 3 consecutive months. Blood was obtained from the abdominal aorta. Serum was isolated. The kidney was obtained. Enzyme linked immunosorbent assay was used to determine 1,25(OH)2D3 contents of renal and serum. Rats were sacrificed at the end of experiment. The thighbone was taken out to determine bone mineral density. The second lumbar vertebra was taken out to measure the expression of lumbar vertebra and renal vitamin D receptor mRNA using real-time fluorescent reverse transcription-polymerase chain reaction. The smal intestine was taken out to measure the expression of smal intestine CaBp-D9K mRNA using real-time fluorescent reverse transcription-polymerase chain reaction. RESULTS AND CONCLUSION:Compared with the sham operation group, bone mineral density of femur, and 1,25(OH)2D3 levels of serum and kidney and the expression of lumbar vertebra vitamin D receptor mRNA significantly decreased in model group, and the expression of smal intestine CaBp-D9K mRNA significantly decreased in model group. Compared with the model group, bone mineral density of femur, and 1,25(OH)2D3 levels of serum and kidney, and the expression of the second lumbar vertebra vitamin D receptor mRNA, and the expression of smal intestine CaBp-D9K mRNA were increased in moderate-and high-dose flavonoids groups and vitamin D3 group. Results indicated that flavonoids from Cuscuta chinensis could significantly raise bone mineral density of femur, and 1,25(OH)2D3 levels of serum and kidney and the expression of lumbar vertebra vitamin D receptor mRNA and the expression of smal intestine CaBp-D9K mRNA, accelerate intestinal calcium absorption and osteoblast activity, and reinforce quality of the bone.

Expression of Notch1, Jagged1 and NICD in epithelial ovarian carcinomas and a preliminary study on the activity of gamma-secretase in epithelial ovarian carcinoma cell lines / 中华妇产科杂志

Objective To study the expression of Notch 1,Jagged1 and Notch intracellular domain (NICD) in epithelial ovarian carcinoma tissues and analyze the clinical significance.To explore the activity of γ-secretase in epithelial ovarian carcinoma cell line SKOV3 and the effect of N-[N-(3,5-dil uorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT),a γ-secretase inhibitor on the activity of γ-secretase in SKOV3.Methods Immunohistochemistry staining method was performed in 43 patients with epithelial ovarian carcinoma and 11 patients with benign epithelial ovarian tumor to detect the expression of Notch1,Jagged1 and NICD.The differences of expressionof Notch1,Jagged1 and NICD between malignant and benign ovarian tumors was compared and alsoanalyzed the correlation with clinicopathological parameters of ovarian carcinoma.Human serous ovarian cancer cell line SKOV3 and immortalized nontumorigenic ovarian epithelial cell line T29 were incubated in vitro.The activities of γ-secretase in SKOV3 and T29 with dimethyl sulfoxide (DMSO) and DAPT were detected respectively by Gal4VP16/UAS and dual luciferase reporter assay system.Results (1) The immunohistochemical composite scores (ICS) of Notch1 in epithelial ovarian carcinoma (6.7±2.2) were not significantly different with those in benign epithelial ovarian tumor (5.4± 2.7,P=0.153),while the ICS of Jagged 1 and NICD in epithelial ovarian carcinoma (5.3± 2.4,5.3± 2.3) were higher than those in benign epithelial ovarian tumor (1.6± 1.4,3.1± 1.7; all P＜0.01).The expression of Notch 1,Jagged 1 and N ICD had no correlation with patients' aged,history of carcinoma,ascites,the level of serum CA125,maximum length of ovarian tumor,Federation International of Gynecology and Obstetrics (FIGO) stage,grade and pathology subtypes (all P＞0.05).The hazard ratio between the high expression of Notch1,Jagged1,or NICD and the moderate to low expression of Notch1,Jagged1,or NICD,and Jagged1 were 0.771,1.648 and 1.316,respectively (all P＞0.05).The 5-year survival rate and median survival time between the high expression of Notch,Jagged 1 or NICD in subgroup and moderate to low expression in subgroup were of no difference (all P＞0.05).The activity of γ-secretase in SKOV3 was significantly higher than that in T29 [(12.2± 1.4)％,P=0.019].(2)After DAPT treated,the relative activity of γ-secretase in SKOV3 (50 μmol/L) was declined from (100.0±5.3)％ to (6.6±0.8)％ (P=0.001).Conclusions Jagged1 and NICD in Notch1 pathway may play a key role in the occurrence of ovarian carcinoma.The activity of γ-secretase in epithelial ovarian carcinoma was higher than that in ovarian epithelial cell which suggest that DAPT,γ-secretase inhibitor,may become the target of ovarian carcinoma treatment.

Clinical research advancement of fecal calprotectin during the neonatal period / 国际儿科学杂志

Neonatal gastrointestinal dysfunction was closely related to postnatal growth retardation.Looking for an early specific indicator to reflect gastrointestinal function was important.Calprotectin was a calciumzinc binding protein mainly in human neutrophil granulocytes,which was the marker of acute inflammatory cell activation.Fecal calprotectin was expressed at high levels during the neonatal period and was significantly higher in the intestinal damage,which could be used as the marker of intestinal illnesses (such as feeding intolerance,neonatal diarrhea,necrotizing enterocolitis).This review will summarize the characteristics of fecal calprotectin in neonates and its clinical significance as a noninvasive marker of intestinal illnesses during the neonatal period.

Implications of calpains in health and diseases.

The number of mammalian calpain protease family members has grown as many as 15 till recent count. Although initially described as a cytosolic protease, calpains have now been found in almost all subcellular locations i.e., from mitochondria to endoplasmic reticulum and from caveolae to Golgi bodies. Importantly, some calpains do not possess the 28 kDa regulatory subunit and have only the 80 kDa catalytic subunit. In some instances, the 80 kDa subunit by itself confers the calpain proteolytic activity. Calpains have been shown to be involved in a number of physiological processes such as cell cycle progression, remodeling of cytoskeletal-cell membrane attachments, signal transduction, gene expression and apoptosis. Recent studies have linked calpain deficiencies or it’s over production with a variety of diseases, such as muscular dystrophies, gastropathy, diabetes, Alzheimer’s and Parkinson’s diseases, atherosclerosis and pulmonary hypertension. Herein, we present a brief overview on some implications of calpains on human health and some diseases.

Effect of estrogen on the expression of matrix GLA protein in ovariectomized SD rats / 中华妇产科杂志

Objective To investigate the effect of estrogen on expression of matrix GLA protein (MGP)in ovariectomized Sprague-Dawley(SD)rats and the role of estrogen in improving postmenopausal osteoporosis.Methods Thirty-six SD female rats were allocated into 3 groups randomly,every 12 rats in ovariectomized group(OVX group),estrogen group(E group)and control group(sham group).Rats in OVX and E group all underwent bilateral ovariectomy,those rats in E group were given by estradiol benzoate intramuscularly after 3 weeks of ovariectomy.Rats in sham group underwent bilateral lipectomy near the ovary.All rats were kept the urine and the serum every three weeks and were sacrificed after 15 weeks.The pathology changes of uterus,lumbar vertebral bones were observed by immunohistochemistry.Bone mineral density(BMD)of lumbar vertebra of rats was determined by dual energy X ray absorptiometry(DEXA).The content of MGP in serum and urine was determined by ELISA.Expression of underearboxylated matrix GLA Protein(MGP)was detected by immunohistochemistry.Relative quantification of MGP mRNA expression in lumbar vertebra bone was detected by Fluorescent real-time quantitative polymerase chain reaction.Results(1)After 15 weeks of ovariectomized,the endometrium of uterus and lumbar vertebra exhibit remarkable pathologic changes in OVX group.The serum estrogen of(454±66)pmol/L in OVX group were lower than in(527 ±77)pmol/L in sham group and(556 ±80)pmol/L in E group significantly (P ＜ 0.05).The BMD of lumbar vertebra of(0.263 ± 0.030)g/cm2 in OVX group were lower than (0.295 ±0.024)g/cm2 in sham group and(0.279 ±0.024)g/cm2 in E group significantly(P ＜0.01).(2)The serum MPG protein in OVX group and E group showed decreased trends after ovariectomized,which were(104 ±64)ng/L in OVX group and(134 ±6)ng/L in E group at 9 weeks,which reached statistical difference(P ＜ 0.05).However,MGP in urine in sham group did not exhibit significant difference after 15 weeks of surgery(P ＞0.05).The MGP in urine of E group showed increased trends after 12 weeks of surgery,which reached(110.0 ±3.4)ng/L at 15 weeks,in the mean time,it was found that(86.5 ±2.5)ng/L of MGP in urine in OVX group,which showed significant difference(P ＜ 0.05).(3)MGP could be observed in lumbar vertebra in OVX group by immunochemistry staining.In the other two groups,the expression of MGP was not dominant.(4)Relative quantification of MGP mRNA expression in lumbar vertebra was defined as 1 in OVX group,when compared with 0.289 ±0.260 in E group and 0.103 ±0.098 in sham group,the difference showed statistically significant(P ＜ 0.01).Conclusion Estrogen could increase the expression of MGP mRNA and protein in ovariectomized rats and might play an important role in improving postmenopausal osteoporosis.