Effect of paracellular permeation enhancers on intestinal permeability of two peptide drugs, enalaprilat and hexarelin, in rats

Transcellular permeation enhancers are known to increase the intestinal permeability of enalaprilat, a 349 Da peptide, but not hexarelin (887 Da). The primary aim of this paper was to investigate if paracellular permeability enhancers affected the intestinal permeation of the two peptides. This was investigated using the rat single-pass intestinal perfusion model with concomitant blood sampling. These luminal compositions included two paracellular permeation enhancers, chitosan (5 mg/mL) and ethylenediaminetetraacetate (EDTA, 1 and 5 mg/mL), as well as low luminal tonicity (100 mOsm) with or without lidocaine. Effects were evaluated by the change in lumen-to-blood permeability of hexarelin and enalaprilat, and the blood-to-lumen clearance of

A Comparison of the Accuracy of Different Single Plasma Sample Methods for Measuring Glomerular Filtration Rate Using ⁵¹Cr-EDTA in Children / 대한핵의학회잡지

PURPOSE: Among the different methods of measuring glomerular filtration rate (GFR) using 51Cr-ethylenediaminetetraacetic acid clearance, the two-plasma-sample method (TPSM) is widely used, and highly accurate. The single-plasma-sample method (SPSM) is occasionally used for simplicity, at the expense of accuracy. Our aims were (1) to investigate the correlation and (2) to compare the accuracy of six known SPSMs in pediatric patients in reference to TPSM.METHODS: We retrospectively reviewed 122 pediatric cases (65 boys, age 7.3 ± 4.6 years) and analyzed 307 GFR measurements. SPSMs included Groth and Aasted at 120 min, Hamat 120 min, Christensen and Groth at 120 and 240 min, and Jacobsson at 120 and 240 min. Reference GFR (GFRref) was defined using TPSM GFR corrected by the Jodal and Brochner-Mortensen equation. GFRref < 30 mL min−1 1.73 m−2 were excluded. The standard error of the estimate (SEE) and the number of cases with differences > 10% (N10%) were used to evaluate accuracy.RESULTS: SPSMs generally correlated well with GFRref (r = 0.92~0.99) and were relatively accurate (SEE = 9.21~15.60). Groth and Aasted showed the smallest SEE, while Jacobsson at 240 min showed the smallest N10% for all GFRref ranges. As for the decreased GFRref, Ham was most accurate followed by Jacobsson at 240 min.CONCLUSIONS: Jacobsson at 240 min provided good accuracy in all GFRref ranges and was well correlated with TPSM. Jacobsson at 240 min might be the most appropriate method to substitute for TPSM in pediatric patients. Ham could be an alternative in patients with impaired renal function.

Effect of genetic modified K562 cells on NK cell markers and function / 中国免疫学杂志

Objective:To study the influence of different culture conditions on charcic and inhibition activity of nature killer cells ( NK) ,whether to join the modified K562 cells with IL-6 cytokine.Methods:According to the 5′end of the human IL-6 cDNA sequence,PCR primers designed to amplificate,express and transfect K562 cells cDNA library as a template for DNA.Genetic modified K562 cells as stimulating cells were prepared by expressing IL-6.To extract peripheral blood mononuclear cells( PBMC) from human peripheral blood.PBMC were explanted by genetic modified K562 stimulated.The expansion was initiated by CO-culture of PBMC and irradiate genetic modified K562 cell.The number of NK cell increased by directed induced generation of genetic modified K562 cell.Immunophenotypic analysis of NK cell surface markers was performed by flow cytometry (FCM).51Cr release assay was employed to measure the specific lysis skilling of NK cell target K562 cells.Results:We have constracted genetic modified K562 cells by genetic engineering.As stimulated cell added into the PBMC,an average of 760 ±18 fold expansion of CD56+CD16+CD3-cells was observed after 3 weeks of co-culture system.The NK cells population could proliferated more 91%±2% after expansion comparing with 6%± 0.4%in PBMC before expansion by FCM.The cytotoxical activity of NK cells which was induced by genetic modified K562 cell was the strongest than induced by IL-6 cytokine alone.The expanded NK cells lysed 92%±2% of K562 targets in a 5∶1 effector to target ratio.In this case,the NK cells induced by genetic modified K562 cells against tumor cells was more lethal.Conclusion:PBMC based in vitro expansion of natural killer cells was set up by genetic modified K562 cells.The cytotoxicity of NK cells was the strongest induced by genetic modified K562 cell treated.These results had important guiding significance for the the NK large number of amplification and used in clinical.

Taxa de filtração glomerular pelo EDTA marcado com 51cromo e com 113míndio em equinos / Glomerular filtration rate by 51chromium and 113m indium labeled EDTA in horses

Determinou-se a taxa de filtração glomerular por meio do EDTA, marcado com 51Cr e com 113mIn, pela técnica de injeção única, em nove equinos, seis machos e três fêmeas com idades de dois a 12 anos. A taxa de filtração glomerular foi calculada a partir da curva de desaparecimento no plasma e do volume de distribuição do radiotraçador, 51Cr-EDTA ou 113mIn-EDTA. O resultado (média±desvio-padrão) foi de 148,80±26,42mL.min-1.100kg. Conclui-se que a medida da taxa de filtração glomerular pelo 51Cr-EDTA ou 113mIn-EDTA, pela técnica de injeção única, elimina o cateterismo vesical e, por sua simplicidade, comodidade, precisão e baixa dose de radiação, pode ser empregada em cavalos como método de escolha em rotina clínica.

The glomerular filtration rate was determined in nine healthy horses, six male and three female, aged two to 12-year-old, by means of 51Cr and 113mIn labeled EDTA single injection technique. The glomerular filtration rate was calculated from the plasma disappearance curve and the volume of distribution of the radiotracer, 51Cr-EDTA or 113mIn-EDTA. The result (mean±standard deviation) was 148.80±26.42mL.min-1.100kg. It is concluded that the measurement of glomerular filtration rate by 51Cr-EDTA or 113mIn-EDTA by single injection technique eliminates the bladder catheterization, and for its simplicity, convenience, accuracy, and low dose of radiation, can be used in horses as a method of choice in clinical routine.

The changes of intestinal permeability in patients with mild acute pancreatitis / 대한내과학회지

BACKGROUND: Many studies in severe acute pancreatitis have demonstrated an increase in intestinal permeability, but not in mild acute pancreatitis. The current methods to measure intestinal permeability need much time and also laborious work. Therefore, we investigate the changes of intestinal permeability in patients with mild acute pancreatis and clinical predictive factor for the intestinal permeability in patients with acute pancreatitis. METHODS: The intestinal permeability were measured in 14 normal heathy controls, 41 patients with mild acute pancreatitis (alcoholic 14, biliary 12, idiopathic 15) by measuring 24 hour urine excretion of 51Cr-EDTA (51Cr-ethylenediaminetetraacetic acid) for evaluation of the gut barrier dysfunction. We compared the intestinal permeability with clinical characteristics of patients. RESULTS: The intestinal permeability was significantly increased in patients with mild acute pancreatitis (6.01+/-4.11%, p<0.001) versus control subjects (1.86+/-0.52%). There was no significant difference in the intestinal permeability among the patients with alcoholic, biliary and idiopathic pancreatitis. The correlation was not found between intestinal permeability and clinical characteristics in patients with mild acute pancreatitis. CONCLUSIONS: The intestinal permeability is increased in patients with mild acute pancreatitis regardless of etiology of pancreatitis. The predictive factor for gut barrier dysfunction is not detected in patients with mild acute pancreatitis.

Expression of human hDAF in CHO cells and its decay-accelerating activity / 第三军医大学学报

Objective To obtain Chinese hamsterovary (CHO) cell line expressing human decay accelerating activity (hDAF) stably and to observe the protective effect of hDAF on heterologous cells under the circumstance of complement activation. Methods The eukaryotic expression vector DAF pcDNA3.1 was constructed and then transfected into CHO cells by lipofection. Monoclones of cells expressing hDAF stably were screened by the method of limiting dilution. hDAF expression was detected by flow cytometry. The decay accelerating activity of hDAF was determined by assay of C3 deposition and 51Cr release. Results The expression vector DAF pcDNA3.1 was successfully constructed, and monoclones of cells expressing hDAF were obtained. CHO cells expressing hDAF could decrease C3 deposition and attenuate the killing effect of activation of the complement system. Conclusion We have obtained CHO cell clones expressing hDAF stably, which is helpful for the further studies of the relationship of the structure with the functions of hDAF.

Estudo do tráfego linfocitário e sua modulação em camundongos, in vivo

Uma compreensão aprofundada dos mecanismos envolvidos na migração linfocitária e na sua modulação, permitiria que essas células fossem direcionadas artificialmente. Isto poderia impedir sua entrada e acúmulo em sítios indesejáveis (como por exemplo, órgãos alvo em doenças auto-imunes ou órgãos transplantados) ou, alternativamente, seria possível facilitar sua em uma situação em que o acúmulo de células efetoras funcionais fosse mandatório, como seria o caso de tumores. No presente trabalho este problema foi abordado estabelecendo-se, inicialmente, um sistema de estudo para padronização da distribuição do trêfego linfocitário, em camundongos normais, e em seguida procurou-se uma possibilidade de modular este tráfego. Foram utilizadas células linfóides marcadas com51 Cr e injetadas por via endovenosa. Duas substâncias, o antiinflamatório não esteróide Indometacina (INDO) e o agente imunossupressor Ciclosporina A (CS-A) foram utilizadas com o objetivo de verificar se seria possível modular esta distribuição linfocitária normal ou em situações inflamatórias desencadeadas por uma reação de sensibilidade de contato e por indução de tumor. CS-A foi capaz de alterar a migração linfocitária, em situações fisiológicas, para o intestino e timo enquanto INDO apenas induziu uma inibição da migração de linfoblastos periféricos para o timo. CS-A aumentou a permeabilidade do timo a linfoblastos periféricos e induziu um maior acúmulo de linfoblastos mesenféricos no intestino. Por outro lado, o tratamento de infiltrados inflamatórios com CS-A produziu efeitos paradoxais. CS-A inibiu o acúmulo de linfoblastos em tumores em pleno crescimento, mas só foi capaz de inibir parcialmente e tardiamente a entrada de células em lesões pelo sesibilizante de contato Oxazolona (OXA). Nas mesmas condições, INDO tem efeito comparável ao de CS-A, nas lesões induzidas por OXA. O tratamento deste infiltrado inflamtorio com ambas, CS-A e INDO não modificaram o efeito exercido por nenhuma delas, considerado isoladamente. O tratamento experimental com radiação, por si mesmo, induziu alterações na migração de células para o baço, LNM, intestino, pele, pulmão e timo. Tal tratamento levou ao acúmulo de linfoblastos no intestino e timo e, surpreendentemente, CS-A produziu um aumento considerável na ligração linfocitária para o intestino e timo de animais irradiados, mostrando um efeito sinergístico, sugerindo que o acúmulo dessas células nos vários modelos resulta de mecanismos diversos e que o uso de CSA não pode ser considerado de maneira exclusiva em inflamatórios de risco de linfócitos.

A better understanding of the mechanism involved on lymphocyte migration and modulation might allow these cells to be artificially directed. This could hinder the entrance and accumulation of these cells at inconvenient sites (target organs in autoimmune diseases or transplanted organs, for example) or alternatively, it woud be possible to facilitade their entrance in situations in which the accumulation of effector functional cells were mandatory as it would be case for tumors. In the present work this problem was dealt with, by initially establishing a system 1· of study to standardize the distribution of the lymphocyte traffic in normal mice and then ; it has been searched a manner of modulating such traffic. Lymphoid cells labeled with 51Cr and intravenously injected have been utilized. Two substances, the anti-inflammatory non steroid - Indometacin (INDO) and the immunosupressor agent - Cyclosporin A (CS-A), were utilized in order to verify the posibility of modulating lymphocyte distribution at inflammatory situations induced by several stimuli. CS-A has been able to alter the lymphocyte migration, in physiological situations, towards gut and thymus whereas INDO only induced an inhibition of the peripheric lymphoblasts migration towards the thymus. CS-A increasing thymus permeability to peripheric Iymphoblasts and induced a higher accumulation of mesenteric lymphoblasts in the gut. On the other hand, treatment of inflammatory infiltrates with CS-A produced paradoxical effects. CS-A inhibited the accumulation of lymphocytes in developing tumors but it was only able to inhibit the entrance of cells in lesions induced by Oxazolone (OXA), partially and at late stage. In the same conditions, INDO has comparable effect as CS-A, in lesions induced by OXA. The treatment of this inflammatory infiltrate with both CS-A and INDO did not modify the effect exerted by either of them, considered isolately. The experimental treatment with radiation, induced, on its own, modifications in the migration of cells towards spleen, mesenteric lymph nodes, gut, lungs and thymus. Such treatment led to the accumulation oflymphoblasts in the gut and surprisingly, CS-A promoted a considerable increase of the lymphocyte migration towards the gut and the thymus of irradiated animals, suggesting that such cell accumulation in the several models results from diverse mechanisms and that the use of CS-A canot be considered in an exclusive way on infiltrates rich with lymphocytes.

The Effects of Ascorbic Acid on Intraocular Pressure Elevation by Viscoelastic Substance

We evaluated intraocular pressure(IOP), remained radioactivity of 51Cr tagged Healon(R) in the anterior chamber following lens aspiration in rabbit eyes, and ultrastructual changes of trabecular meshwork were observed after intravenous injection of ascorbic acid. The average lOPs returned to preoperative lOP level on the postoperative seventh day in control group. While, those in experimental group returned on the postoperative third day. Healon(R) was completely outflowed in control group on the postoperative seventh day and in experimental group, on the postoperative third day. There is a significant correlation between lOP and remained radioactivity rate of Healon(R) in control and experimental group. Under scanning electron microscope examination, the control group showed the most of Healon(R) debris outflowed on the postoperative seventh day. On the other hand, the experimental group showed no Healon(R) debris was observed on the postoperative third day. In conclusion, our experiment showed ascorbic acid is effective in lowering intraocular pressure and preventing chance of intraocular pressure elevation.

The influence of anisodamine on the cerebral blood flow in uncompletely ischemic rat brains / 中国药理学通报

The influence of anisodamine on the cerebral blood flow in the uncompletely ischemic brains was studied with radioactive biomicrosphere assay. After the bilateral carotid arteries were occluded the cerebral blood flow drawmatically decreased and middle part of the hemisphere had the most severe reduction. Anisodamine 10mg ? kg-1 iv selectively increased the cerebral blood flow in this area and 20mg ? kg-1 increased that in the whole hemisphere. When it was used in the dose of 40mg ? kg-, the increasing effect on the cerebral blood flow became weaker. While the cerebral blood flow decreased, the myocardial blood flow increased significantly after bilateral carotid artery occlusion. Anisodamine 20mg ? kg-1 iv made it further increase and 40mg ? kg-1 iv made it decrease. The cardiac index was not influenced by bilateral carotid artery occlusion and anisodamine 10~20mg ? kg-1 iv, but was significanntly increased by anisodamine 40mg ? kg-1 iv. These results suggested that anisodamine may be used in the treatment of ischemic cerebral diseases and should be used in different doses according to different purposes.

EFFECTS OF METRONSDAZOLE DERIVATIVE ON MEMBRANE PERMEABILITY OF~51Cr LABELLED L_7712 CELLS / 第二军医大学学报

This report describes the effects of a new radiosensitizer, metronidazole derivative (CM) synthesized in our laboratory, on membrane permeability of 51Cr labelled L7712 cells.It was found that CM could be incorporated into the cancer cells easily, and resulted in increased membrane permeability.Under aerobic condition, the release rate of 51Cr was increased when the cells were irradiated with more than 40 Gy Y-rays.At a dose of 0-80 Gy, the sensitization effects of CM on membrane permeability of irradiated cells were not found.

Studies on T cell,T Cell Subpopulation and Natural Cell Activities of the Peripheral Blood Lymophocytes from Patients with Tumor / 中国免疫学杂志

In this paper,we reported that T cell,T cell subpopulation and natural killer(NK)cell activities of peripheral blood lymophocytes from the Fatients with tumor weredetected with OKT monoclonal antibody and ~(51)Cr release cytotoxity test.The resultsshowed that percentage of OKT3~+,OKT4~+ cell in the peripheral blood of patients withtumor are low,OKT8~+ is high and the ratio of OKT4/OKT8 is low.The results haveproved that the ratio of OKT4/OKT8 and NK cell activities were significantly low inthe peripheral blood of patients with tumor.However,the activities have restored tonormal or near normal level in the patients after surgical operation for more that twoyears with no recurrence. Our data suggest that detecting of the ratio of OKT4/OKT8 and activities of NKcell might be valuable to the prognosis of patients with tumor.