Synovium and blood capillaries at the femoral attachment of anterior cruciate ligament are significantly abundant than those at the tibial attachment / Sinovial y capilares sanguíneos en la inserción femoral del ligamento cruzado anterior son significativamente más abundantes que los de la inserción tibial

SUMMARY: The anterior cruciate ligament (ACL) is a ligament that mainly controls the anterior and rotational mobility of the knee joint, and its surface is covered by a synovial membrane with large number of blood vessels. In general, nutritional supply to the ligament is from many capillaries in the adjacent synovium. However, statistical studies of the capillaries distributed to the ACL are insufficient. In this study, we examined cross-sectional histological images of the femoral attachment (femoral level), middle level of the tendon (middle level), and tibial attachment (tibial level) of the ACL and statistically analyzed blood capillary distribution among the three levels. The ACLs of 10 cadavers were divided into 5 equal sections, and 4mm-thick paraffin sections were made at the femoral level, middle level, and tibial level, and then hematoxylin-eosin (HE) staining were performed. The area of each transverse section was measured using Image-J 1.51n (U. S. National Institutes of Health, Bethesda, MD, USA). Fiber bundles of the ACL were relatively small and sparse in cross-sectional area at the femoral level and became larger and denser toward the tibial level. Many blood levels. The synovium at the attachment of ACL covered the surface of the fiber bundle and also penetrated deeply between the fiber bundles. In particular, the blood capillaries were densely distributed in the synovium at the femoral attachment rather than another two levels. Indeed, the number of capillaries were also most abundant in the femoral level. The cross-sectional ACL area at the femoral level is significantly small, however, the blood capillaries were most abundant. Therefore, when the ACL is injured, its reconstruction with preservation of the femoral ligamentous remnant may be clinically useful for remodeling of the grafted tendon.

El ligamento cruzado anterior (LCA) es un ligamento que controla principalmente la movilidad anterior y rotacional de la articulación de la rodilla, y su superficie está cubierta por una membrana sinovial con gran cantidad de vasos sanguíneos. En general, el suministro de nutrientes al ligamento proviene de muchos capilares en la sinovial adyacente. Sin embargo, los estudios estadísticos de los capilares distribuidos en el LCA son insuficientes. En este estudio, examinamos imágenes histológicas trans- versales de la inserción femoral (nivel femoral), el nivel medio del tendón (nivel medio) y la inserción tibial (nivel tibial) del LCA y analizamos estadísticamente la distribución de los capilares sanguíneos entre los tres niveles. Los LCA de 10 cadáveres se dividieron en 5 secciones iguales y se realizaron cortes en parafina de 4 µm de espesor a nivel femoral, medio y tibial, y luego se realizó tinción con hematoxilina-eosina (HE). El área de cada sección transversal se midió utilizando Image-J 1.51n (Institutos Nacionales de Salud de EE. UU., Bethesda, MD, EE. UU.). Los haces de fibras del LCA eran relativamente pequeños y escasos en el área de la sección transversal a nivel femoral y se hicieron más grandes y más densos hacia el nivel tibial. La membrana sinovial en la unión del LCA cubría la superficie del haz de fibras y también penetraba profundamente entre entre los haces de fibras. En particular, los capilares sanguíneos estaban densamente distribuidos en la unión femoral de la sinovial respecto a los otros dos niveles. De hecho, el número de capilares también fue más abundante a nivel femoral. El área transversal del LCA a nivel femoral era significativamente pequeña, sin embargo, los capilares sanguíneos fueron los más abundantes. Por lo tanto, cuando hay una lesión del LCA su reconstrucción con preservación del ligamento femoral remanente puede ser clínicamente útil para remodelar el tendón injertado.

Role of M3 receptor in penehyclidine hydrochloride-induced reduction of increased permeability of human pulmonary microvascular endothelial cells caused by endotoxin: the relationship with MAPK signaling pathway / 中华麻醉学杂志

Objective To evaluate the role of M3 receptor in penehyclidine hydrochloride(PHC)-induced reduction of increased permeability of human pulmonary microvascular endothelial cells (PMVECs) caused by endotoxin and the relationship with mitogen-activated protein kinase (MAPK) signaling pathway.Methods Human PMVECs were seeded in 6-well plates (2 ml/hole) or in culture flasks (4 ml/flask) at the density of 1 × 105 cells/ml and randomly divided into 6 groups (n=5 each):control group (group C),M3 receptor shRNA transfection group (group shRNA),lipopolysaccharide (LPS) group,penehyclidine plus LPS group (group P+LPS),LPS plus M3 receptor shRNA transfection group (group LPS+shRNA) and PHC plus LPS plus M3 shRNA transfection group (group P+LPS+shRNA).The cells were transfected with shRNA plasmid containing 2.5 nmol/L M3 receptors in shRNA,LPS+shRNA and P+LPS+shRNA groups.LPS at the final concentration of 0.1 μg/ml was added at 24 h of incubation and then cells were incubated for 1 h in LPS and LPS+shRNA groups.PHC at the final concentration of 2 μg/ml was added at 24 h of incubation,cells were incubated for 1 h,then LPS at the final concentration of 0.1 μg/ml was added,and cells were incubated for another l h in P+LPS and P+LPS+shRNA groups.The permeability of PMVECs was measured using Transwell assay.The expression of phosphorylated p38 MAPK (p-p38 MAPK)and phosphorylated extracellular signal-regulated protein kinase 1/2 (p-ERK1/2) was detected by Western blot,the expression of heat shock protein 27 (HSP27) using immunofluorescent staining,and the expression of M3receptor mRNA by real-time polymerase chain reaction.Results Compared with group C,M3 receptor mRNA expression was significantly down-regulated in group shRNA,and the permeability of cells was significantly increased,and the expression of p-p38 MAPK,p-ERK1/2,HSP27 and M3 receptor mRNA was up-regulated in group LPS (P＜0.05).The permeability of cells was significantly decreased,and the expression of p-p38 MAPK,p-ERK1/2,HSP27 and M3 receptor mRNA was down-regulated in P+ LPS,LPS+shRNA and P+LPS+shRNA groups as compared with group LPS,and in group P+LPS+shRNA as compared with group LPS+shRNA (P＜0.05).Conclusion The mechanism by which PHC reduces endotoxin-caused increased permeability of human PMVECs is related to inhibiting activation of MAPK signaling pathway after down-regulating M3 receptor.

Effect of Compound Nitroglycerin Gel on Skin Ulcer Wound in Rats / 医药导报

Objective To study the protective effect of compound nitroglycerin gel on rats with skin ulcer wound and its action mechanism. Methods Skin ulcer modle of 54 rats was established.Then the rats were randomly divided into three groups (n=18 each), including model control group, Jin wan hong group, and compound nitroglycerin gel group. Wound healing process and healing time were recorded.At the day 7 and 14 after the model was established, the number of fibroblasts and new blood capillaries of granulation tissue from center of the wound were measured,and RT-PCR was used to detect mRNA expression levels of VEGF, Ang1 andHIF-1α. Results As compared with model control group [(24.17±5.91) days], healing time of skin in the compound nitroglycerin gel group was significantly shorter [(14.67±3.76) days, P<0.01].The numbers of fibroblasts (61.20±7.56) and new blood capillaries (9.35±1.43) were increased, and mRNA expression levels of VEGF (1.692±0.196), HIF-1α (1.527±0.174) and Ang1 (1.548±0.203) were remarkably up-regulated on 7th day (P<0.05 or P<0.01).While on 14th day, the numbers of fibroblasts (28.00±5.96) and new blood capillaries (4.20±1.30) were decreased and the mRNA expression levels of VEGF (1.156±0.123), HIF-1α(1.021±0.105) and Ang1 (1.034±0.134) were significantly down-regulated (P<0.05 or P<0.01) . Conclusion Compound nitroglycerin gel can treat skin ulcer wound via regulating the numbers of fibroblasts, new blood capillaries and VEGF/Ang1/HIF-1α signal transduction pathway.

Desarrollo y evaluación de un protocolo serológico de fluorescencia polarizada en el estudio de anticuerpos contra Brucella spp en humanos / Development and evaluation of a serological protocol of fluorescence polarization for the preliminary study of Brucella spp antibodies in humans

A objeto de mostrar el desarrollo y alcance de un método de análisis serológico basado en la técnica de fluorescencia polarizada (FPA) a partir de una gota de sangre obtenida mediante punción capilar, se realizó la determinación de anticuerpos antibrucelosis de un conjunto de 321 personas de alto riesgo laboral. Los resultados se compararon con la data proveniente del análisis de sueros sanguíneos mediante FPA e inmunoanálisis enzimático competitivo (ELISA-c). El número de concordantes fue 318 (99,06%), los 3 discordantes (0,93%) resultaron negativos con fluorescencia polarizada en suero (FPAs) y ELISA-c, pero positivos con FPA capilar (FPAc). Los resultados comparativos de FPAc fueron: sensibilidad: 100%; especificidad: 99,05%; valor predictivo positivo: 66,67%; valor predictivo negativo: 100,0%; proporción de falsos positivos: 0,95%; proporción de falsos negativos: 0%; exactitud: 98,0%; razón de probabilidades: 203,00. La J de Youden para ambos métodos de FPA fue de 0,667. La determinación se consideró confiable y la concordancia de ambos procedimientos de FPA y ELISA-c resultó sin diferencias estadísticas (P>0,05%), lo que permite recomendar ampliamente la implementación del estudio de la brucelosis humana con sangre proveniente de punción capilar como método preliminar.

In order to show the development and scope of a serological analysis method based on fluorescence polarization assay (FPA) from a drop of blood obtained by the capillary technique, a Brucella antibody assay was performed on a group of 321 high-risk workers. The results were compared with data from the analysis of blood serum by FPA and a competitive enzyme immunoassay (ELISA-c). The number of concordance was 318 (99.06%), and discordant 3 (0.93%), which were negative in serum by fluorescence polarization (FPAs) and ELISA-c, but positive with capillary FPA (FPAc). The comparative results FPAc were: sensitivity 100%; specificity: 99.05%; positive predictive value 66.67%; negative predictive value 100.0%; false positive rate: 0.95%; false negative rate: 0%; accuracy: 98.0%; odds ratio: 203.00. The youden J for both FPA methods was 0.667. The identification was considered reliable and the correlation of both procedures, FPA and ELISA-c, was no statistically different (P> 0.05%), which allows to highly recommend the study implementation of human brucellosis with capillary blood as a preliminary method.

Study on Vasodilating Mechanism of the Acetone Extract from Cortex Mori / 中药新药与临床药理

Objective To study the vasodilating effect of the acetone- extract (AE) from Cortex Mori (CM) and its vasodilating mechanism. Methods Guinea pig models with contraction of mesentery blood capillary induced by noradrenalin were used to study the in- vivo vasodilating effects of AE from CM. In- vitro effect of AE from CM on rat thoracic aortic ring was observed. The contents of nitric oxide (NO), nitric oxide synthase (NOS), constitutive NOS(cNOS) and inducible (iNOS) in plasma and aortic tissue were determined by method of nitroreductase chromatometry. Results AE from CM had vasodilating effect on the contracti on of mesentery blood capillary in guinea pigs and the contraction of rat thoracic aortic ring induced by 10? mol/L phenylephrine, even under the condition of pre- treating with glibenclamide (1 ? mol/L) or propranolol (3 ? mol/L). When the endothelium of the rat thoracic aortic ring was removed, AE from CM had not vasodilating effect but constrictive effect. AE from CM could also increase the contents of NO, NOS and cNOS in the aortic tissue and had no obvious effect on the NO content in plasma and iNOS in aortic tissue of rats. Conclusion The vasodilating mechanism of AE from CM may be related to promoting the release of NO from endothelium of blood vessel and synthesis of NOS and cNOS in aortic tissue.