Effect of Asiaticoside on Expression of Th17/Treg Cells in DBA/1 Mice with Collagen-Induced Arthritis / 中国实验方剂学杂志

ObjectiveTo observe the effect of asiaticoside （AC） on the expression of T helper 17 （Th17） cells and regulatory T （Treg） cells in DBA/1 mice with collagen-induced arthritis （CIA）. MethodMale SPF DBA/1 mice were randomized into six groups according to body weight： control group， CIA group， methotrexate group （MTX group， ip， 0.5 mg·kg-1）， and AC low-， medium-， and high-dose groups （ig， 5， 15， 45 mg·kg-1， respectively）. Modeling was performed in rats other than the control group. To be specific， they were immunized with bovine type Ⅱ collagen and complete Freund's adjuvant on the first day and with bovine type Ⅱ collagen and incomplete Freund's adjuvant on the 21st day. Administration began on the day of the second immunization， once a day for 28 days. On the 49th day， related tissues were collected. Then， hematoxylin-eosin （HE） staining was performed to observe the pathological changes of the joints. Immunohistochemical method was used to detect the expression of interleukin-17 （IL-17） and forkhead box protein-3 （FoxP3）， the markers of Th17 and Treg cells， respectively， immunofluorescence double staining the expression of IL-17 and FoxP3 in CD4+T cells of mouse joint tissue， and flow cytometry the proportions of Th17 and Treg cells in mouse lymph nodes. ResultCompared with the control group， CIA group demonstrated joint disorder， damage of articular cartilage and bone， severe bone erosion （P<0.01）， increase in stained CD4 and IL-17 and the integral absorbance （IA） （P<0.01）， decrease in stained FoxP3 and the IA （P<0.01）， rise of Th17/Treg ratio （P<0.01）， elevation of Th17 expression in mouse lymph nodes （P<0.01）， and reduction in Treg expression （P<0.01）. Compared with CIA group， MTX group and three AC groups showed normal joints， alleviated bone erosion and damage， intact and smooth joint surface， and decrease in stained IL-17 and IA （P<0.05， P<0.01）， and MTX group and AC medium-dose and high-dose groups registered decrease in stained CD4 and IA （P<0.01） and reduction in Th17/Treg ratio （P<0.05， P<0.01）. Moreover， AC medium-dose and high-dose groups showed rise in stained FoxP3 and IA （P<0.05， P<0.01）. In the lymph nodes of mice， decrease in expression of Th17 cells （P<0.05， P<0.01） and the increase in expression of Treg cells （P<0.05， P<0.01） were observed in all the three AC group. ConclusionAC can regulate Th17/Treg balance by inhibiting the expression of Th17 cells and promoting the expression of Treg cells in CIA mice.

Effect of Fangji Huangqitang on DBA/1 Mice Collagen Induced Arthritis and Synovial Angiogenesis / 中国实验方剂学杂志

Objective:To observe the effect of Fangji Huangqitang （FJHQT） on collagen induced arthritis （CIA） and synovial angiogenesis in DBA/1 mice. Method:DBA/1 mice were randomly divided into normal group， CIA group and FJHQT group. DBA/1 mice in CIA group and FJHQT group were immunized with bovine type Ⅱ collagen and complete Freund's adjuvant on the first day， and DBA/1 mice were immunized with bovine type Ⅱ collagen and incomplete Freund's adjuvant on the 21^st day to establish CIA model. On the day of the second immunization， the drug was given by gavage once a day for 28 days. On the 22^nd day， the arthritis score and other symptoms of CIA mice were observed. On the 49^th day， Hematoxylin eosin （HE） staining was carried out to observe the angiogenesis in the synovium of CIA mice， the expression of vascular endothelial cell marker platelet endothelial cell adhesion molecule-1 （CD31） and vascular endothelial growth factor （VEGF） in the synovium of CIA mice were detected. Immunofluorescence double staining was used to detect the mature and immature vessels in the synovium of CIA mice. And the microvascular growth of the rat thoracic aortic ring was induced by VEGF （20 μg·L^-1）. The effects of FJHQT （0.25， 0.5， 1 g·L^-1） at different concentrations were observed under microscope. Result:Compared with the normal group， the inflammation， joints， red and swelling of the inflammatory joints of the CIA group were significantly increased （<italic>P</italic><0.01）. The scores of clinical arthritis， the incidence rate， synovial inflammation and angiogenesis were significantly increased （<italic>P</italic><0.01）. The density of blood vessels， the positive expression of CD31 and VEGF， the number of immature vessels in synovial membrane were significantly increased （<italic>P</italic><0.01）. And compared with the CIA group， the inflammation， joint swelling， and malformation of the FJHQT group were significantly improved， the clinical arthritis score， incidence rate， synovial inflammation and angiogenesis were significantly reduced （<italic>P</italic><0.01）. The vascular density， the positive expression of CD31 and VEGF， and the number of immature blood vessels in synovial membrane were significantly increased （<italic>P</italic><0.01）. Compared with blank group， VEGF could significantly induce the growth of microvasculature in rat thoracic aortic ring （<italic>P</italic><0.01）. Compared with VEGF group， FJHQT（0.25， 0.5， 1 g·L^-1） could significantly inhibit the formation of microvasculature in rat thoracic aortic ring （<italic>P</italic><0.01）. Conclusion:FJHQT can effectively alleviate the clinical symptoms and condition of CIA mice， reduce the clinical arthritis score and incidence rate，and inhibit the synovial angiogenesis of CIA mice joints and VEGF induced microvascular formation in rat thoracic aortic rings.

A newly validated high-performance liquid chromatography method with diode array ultraviolet detection for analysis of the antimalarial drug primaquine in the blood plasma

Abstract INTRODUCTION: Primaquine (PQ) diphosphate is an 8-aminoquinoline antimalarial drug with unique therapeutic properties. It is the only drug that prevents relapses of Plasmodium vivax or Plasmodium ovale infections. In this study, a fast, sensitive, cost-effective, and robust method for the extraction and high-performance liquid chromatography with diode array ultraviolet detection (HPLC-DAD-UV ) analysis of PQ in the blood plasma was developed and validated. METHODS: After plasma protein precipitation, PQ was obtained by liquid-liquid extraction and analyzed by HPLC-DAD-UV with a modified-silica cyanopropyl column (250mm × 4.6mm i.d. × 5μm) as the stationary phase and a mixture of acetonitrile and 10mM ammonium acetate buffer (pH = 3.80) (45:55) as the mobile phase. The flow rate was 1.0mL·min-1, the oven temperature was 50OC, and absorbance was measured at 264nm. The method was validated for linearity, intra-day and inter-day precision, accuracy, recovery, and robustness. The detection (LOD) and quantification (LOQ) limits were 1.0 and 3.5ng·mL-1, respectively. The method was used to analyze the plasma of female DBA-2 mice treated with 20mg.kg-1 (oral) PQ diphosphate. RESULTS: By combining a simple, low-cost extraction procedure with a sensitive, precise, accurate, and robust method, it was possible to analyze PQ in small volumes of plasma. The new method presents lower LOD and LOQ limits and requires a shorter analysis time and smaller plasma volumes than those of previously reported HPLC methods with DAD-UV detection. CONCLUSIONS: The new validated method is suitable for kinetic studies of PQ in small rodents, including mouse models for the study of malaria.

Lowing-intraocular pressure and improving aqueous humor dynamic effects of qingguangan drug granules in spontaneous glaucoma mice / 中华实验眼科杂志

Background Researches showed that the increase of intraocular pressure (IOP) in glaucomatous eye is associated with the increasing resistance to aqueous humor outflow effects of transforming growth factor-β (TGF-β) and CD44.Qingguangan is a traditional Chinese medicine and used to treat glaucoma.However,its mechanism of lowing-IOP effect is not elucidated.Objective This study was to investigate the lowing-IOP effect and mechanism of qingguangan granule in DBA/2J mouse,a spontaneous glaucoma model mice.Methods Ten 3 month-old female DBA/2J mice with normal IOP were chosen as control group,and 20 spontaneous ocular hypertension mice aged 9 months were randomized into high IOP group and qingguangan-treated group,with 10 mice for each group.The qingguangan (2.5 g/kg) was administered by gavaging twice per day for consecutive 15 days in the qingguangan-treated group,and normal saline solution was used in the same way in the control group and high IOP group.IOP was measured by anterior chamber injection/suction system at a perfusion rate of 2.5 and 5.0 μl/min,respectively,and the coefficient of aqueous outflow facility (C value) and outflow resistance (R value) were calculated.Another 60 3-month-old DBA/2J mice were randomized into blank control group gavaged with normal saline solution and high-,middle-and low-dose qingguangan groups gavaged with 25.00,12.50 and 6.25 g/kg drugs,respectively,and the mouse serum containing drugs was extracted 7 days after treatment.The scleral tissue with trabecular meshwork were obtained for the culture of trabecular meshwork cells and the cells were identified by immunohistochemistry of fibronectin (FN),laminin (LN) and neuronspecific enolase (NSE).TGF-β was added into the medium for 24 hours with the final concentration of 0,5,10,20,50 and 100 ng/ml,and MMT chromatometry was employed to detect the cell vitality.The cells pre-treated with 20 ng/ml TGF-β were treated with different concentration of drug serum for 24,48 and 72 hours,and the level of TGF-β2 receptor in cell supernatant and the expression of CD44 protein in the cells were detected by ELISA and Western blot assay,respectively.Results The IOPs with perfusion both 2.5 μl/min and 5.0μl/min in the qingguangan-treated group and the control group were significantly lower than those in the high IOP group (all at P＜0.01).Compared with the high IOP group,the C value was significantly reduced (2.35±1.34 vs.1.08±0.36) and the R value was evidently elevated (0.64±0.55 vs.1.05± 0.47) in the qingguangan-treated group (all at P＜0.01).Cultured cells were spindle-shaped with the positive response to FN,LN and NSE antibody.The cell vitality was lower in the 5,10 and 20 ng/ml TGF-β group than that in the 0 ng/ml TGF-β group (all at P＜0.05).Compared with the blank control group,the TGF-β2 receptor content in the supernatant and the related expression level of CD44 protein in the cells were elevated in the TGF-β-treated group (all at P＜0.01),and TGF-β2 receptor contents and CD44 expression levels in the TGF-β+high dose drug serum group was significantly lower than those in the TGF-β group and TGF-β +low dose drug serum group 24,48 and 72 hours after culture (all at P＜0.01).Conclusions Qingguangan can lower IOP of spontaneous glaucoma mice by affecting aqueous humor dynamics.Serum containing qingguangan down-regulates the expressions of TGF-β2 receptors and CD44 in trabecular meshwork cells in vitro.

Immunization with mixed peptides derived from glucose-6-phosphate isomerase induces rheumatoid arthritis in DBA/1 mice / 中国病理生理杂志

[ ABSTRACT] AIM:To establish an animal model of rheumatoid arthritis ( RA) in DBA/1 mice induced by im-munodominant mixed peptides derived from glucose-6-phosphate isomerase (GPI).METHODS: The DBA/1 mice were immunized with emulsified mixed peptide fragments of hGPI 325-339+hGPI469-483 or single peptide hGPI325-339 in com-plete Freund′s adjuvant by subcutaneous injection to induce the model of RA .Body weight , ankle joint symptom scores , the pathological change of the ankle joint , the levels of CD4 +T cells in the spleen and peripheral blood , the proportion of iNKT cells in the peripheral blood , and the levels of TNF-αand IL-6 in serum were detected to evaluate and analyze the model.RESULTS:The hind paw of the model mice appeared red swelling on the 8th day, and then aggravated gradually to the limbs.The red swelling reached peak on the 14th day, and then relieved gradually .Inflammation response dominated by lymphocytes and monocytes was observed in the ankle joint .The inflammatory effect of mixed peptides was more obvious than that of the single one (P<0.05).Compared with control group and the mice treated with single peptide , the weight gain was slow, the amount of CD4 +T cells in the peripheral blood and spleen were increased , the proportion of peripheral iNKT cells in the inflammatory peak was decreased (P<0.05), and the serum level of TNF-αwas increased significantly ( P<0.05) in the mice treated with mixed peptide fragments .CONCLUSION: The immunological characteristics of RA model induced by mixed GPI peptides in DBA/1 mice is closer to that in RA patients , especially in the immunopathology of iNKT cells.Therefore, this model can be used as a new tool for studying the mechanism and immunological intervention of RA.

Estudo da alteração do esvaziamento gástrico em roedores

Fatores que alteram os níveis plasmáticos de substâncias químicas e, por conseguinte, modificam a sua cinética, como por exemplo, a gravidez, podem ter impactos sobre a segurança e eficácia de medicamentos. Em estudo recente, realizado por Carmo (2015), no Laboratório de Toxicologia Ambiental do Departamento de Biologia da Escola Nacional de Saúde Pública da Fundação Oswaldo Cruz (ENSP/FIOCRUZ), foi observado que a concentração plasmática do antimalárico difosfato de primaquina em camundongos fêmeas grávidas DBA/2 era menor do que a concentração do fármaco registrada em igual intervalo de tempo pós-adminstração em camundongos fêmeas não grávidas. Vários estudos sugerem que a diminuição da concentração plasmática de fármacos na gestante pode se dever a um retardo no esvaziamento gástrico e/ou um aumento no volume de distribuição. Alterações do trânsito no trato gastrintestinal podem influenciar diretamente a absorção de fármacos, resultando em absorção mais rápida ou mais lenta. O fármaco analgésico e antipirético paracetamol é absorvido quase que exclusivamente no intestino. Assim a velocidade da sua absorção depende do tempo de esvaziamento gástrico. Fatores tais como alimentação, idade, gravidez e/ou o uso de fármacos que promovem aceleração (metoclopramida) ou o retardo (morfina) da motilidade gastrointestinal, podem influenciar em sua absorção. O objetivo desse trabalho foi desenvolver e padronizar uma metodologia de análise do paracetamol que permitisse investigar o efeito da gravidez sobre o esvaziamento gástrico sobre a cinética de fármacos administrados em pequenos roedores. O método empregado para determinar as concentrações plasmáticas de paracetamol foi a cromatografia em fase líquida de alta eficiência com detector por arranjo de diodos e visualização no ultravioleta (CLAE-DAD-UV), em equipamento Shimadzu Class-VP...

Factors that affect plasma levels of chemicals, and consequently their kinetics, such as pregnancy, can impact on the safety and efficacy of medicines. In a recent study, conducted by Carmo (2015) at the laboratory of Environmental Toxicology (Department of Biological Sciences, National School Public Health, Oswaldo Cruz Foundation -ENSP / FIOCRUZ), it was shown that plasma concentrations of the anti-malarial drug primaquine diphosphate in pregnant female DBA/2 mice were lower than levels found in non pregnant female mice. During pregnancy a delayed gastric emptying and/or an increased volume of distribution may result in lower drug plasma concentrations. Pregnancy-produced changes in the gastrointestinal transit may influence drug absorption. Depending on whether gastric emptying is accelerated or slowed and on the place where drug absorption takes place (stomach or intestines) absorption can be accelerated or slowed. Paracetamol, an analgesic and antipyretic drug, is absorbed almost exclusively in the intestines and is used to investigate the effects of treatment on the gastric emptying rate. Factors such as diet, age, pregnancy or the administration of drugs which accelerate (metoclopramide) or delay (morphine) gastric emptying influence the absorption of paracetamol. The aim of this study was to develop and standardize a methodology to investigate the effect of gastric emptying on the kinetics of drugs administered in small rodents. The methodology used in the analysis of plasma concentrations of paracetamol was High Performance Liquid Chromatography coupled to diode-array detector and visualization on ultraviolet range (HPLC-DAD-UV), using a Shimadzu Class-VP equipment...

Diamond Blackfan Anaemia with Hypothyroidism and CNS Malformations.

Diamond Black fan Anemia (DBA) is a congenital erythroid aplasia that usually presents in infancy. The DBA patients have low red blood cell count (Anaemia). The rest of their blood cells (Platelets & WBCs) are normal. We present a 14 month old male child who presented with severe anaemia. The patient was transfusion dependent since 4 months of age. Clinical examination revealed delayed mile stones and a couple of congenital deformities. Haematological parameters showed elevated foetal haemoglobin level (Hb F – 11.8% ) and elevated serum TSH (thyroid stimulating hormone) level. Peripheral blood picture showed gross microcytic hypochromic red blood cells and absence of reticulocytes with normal levels of leucocytes and platelets. A bone marrow showed gross suppression of Erythroid series with M:E ratio of 30:1. Some large pronormoblasts were found. Family history was not significant. Compiling the clinical features, haematological parameters, PS and bone marrow findings, a diagnosis of DBA was given.

Light Microscopic and Immunohistochemical Study Revealed Sex-Dependent Kidney Morphology in DBA/2crSlc Mice.

Aim: Sex-dependent differences in kidney histology have been observed in different species of the laboratory animals. The present study was conducted to evaluate the sex and strain-dependent changes in DBA/2CrSlc mouse kidney morphology by using light microscopy and immunohistochemistry. Methods: A total of 12 DBA/2CrSlc male and female mice of 2 months of age were used in this study. Mice were sacrificed by exsanguination under anesthesia using a mixture of Ketamine and Medetomidine. Both right and left kidneys were removed aseptically and central slices including hilum were cut perpendicular to the long axis of the organ and preserved in Zamboni solution. Paraffin blocks were made and tissue sections were stained with hematoxylin and eosin, and PAS stains to observe the general morphology of the kidney glomerulus. Immunohistochemistry was performed to detect renin positive sites, expression of Cyclooxygenase-2 (COX-2) and Nitric Oxide Synthase (nNOS). Number of renin, COX-2 and nNOS positive sites were counted and tabulated. The data were statistically analyzed for any significant differences between male and female mice. Results: Our results reveal that the glomerular capsule of male mouse kidney was consisted of a single layer of simple cuboidal epithelium whereas it was a single layer of simple squamous epithelium in the female kidney. PAS-positive granules (small and giant granules) were observed in PST epithelium and collecting ducts in female kidney, but this feature was absent in male kidneys. Strong nNOS positive reaction for PST epithelium and collecting ducts was observed in female, but this character was absent in male kidney. The total number of glomeruli, renin, COX-2, and nNOS positive sites was comparatively higher in female kidneys then that in male. However, statistical analysis revealed no significant differences of the areas of renin, nNOS and COX-2-positive sites between the male and female kidneys (P<0.05). Conclusion: Light microscopic and immunohistochemical study revealed sex-dependent histological morphology of the DBA/2CrSlc mouse kidney. DBA/2CrSlc female mouse kidney revealed renin, COX-2 and nNOS -positive reactions in the present study but male mice showed nNOS-negative reaction. The reason for nNOS-negative reaction in male is not clearly understood. It is suggested that this species can be experimentally used in the laboratory for investigating kidney function and related pathological studies.

All-Trans Retinoic Acid Attenuates Interleukin-23/Interleukin-17 Pathway and Promotes Skin Allograft Survival in Mice / 天津医药

Objective To investigate the effects of all-trans retinoic acid (ATRA)-intragastric-administration on the survival time of mouse skin allografts and the role of interleukin (IL)-23 and IL-17 thereof. Methods The skin trans-plantation of mice was done by DBA/2 as donors and Balb/c as recipients. The recipients were divided randomly into three groups:control group, low-dose group and high-dose group. Mice of the corresponding groups were intragastrically adminis-tered corn oil, 10 mg/kg ATRA and 30 mg/kg ATRA respectively from 1 day before the transplantation to the 14th day after the transplantation. The survival time of transplanted skin was observed after the operations. Skin grafts of mice were harvested for histopathological examination in three groups. The serum levels of IL-23 and IL-17 were measured by enzyme-linked im-munosorbent assay (ELISA). The expression levels of IL-23, RORγt and IL-17 mRNA in skin allografts were detected by re-al-time fluorescent quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Results Compared with con-trol group, the average survival time of mouse skin allografts was significantly prolonged in low-dose group and high-dose group (P＜0.05). The less lymphocyte infiltration and destruction of architecture were found in the skin biopsies. The serum expression of IL-23 protein was lower (P＜0.05), but no significant difference was found in two treatment groups. The serum expression levels of IL-17 protein were reduced in turn in receptors of control group, low-dose group and high-dose group (P ＜ 0.05). The expression levels of IL-23, RORγt and IL-17 mRNA in skin grafts were significantly lower in low-dose group and high-dose group than those of control group (P＜0.05), but no significant difference was found in two treatment groups. Conclusion ATRA can effectively prolong the survival time of skin allografts, which may related with the inhibi-tion of the expression of IL-23, RORγt and IL-17 mRNA and the development of IL-23 and IL-17 protein.

Evaluation of the in vitro and in vivo anticancer properties of Moroccan propolis extracts

This investigation aimed to evaluate the in vitro and in vivo antitumor potential of a Moroccan propolis extracts. For in vitro assays, three mammalian tumor cell lines were used: BSR (hamster renal adenocarcinoma), Hep-2 (human laryngeal carcinoma) and P815 (murin mastocytoma). The propolis ethanolic extract as well as the ethyl acetate extract, exert an in vitro cytotoxic activity in dose-dependent manner. The IC50 values were ranging from 15 µg/mL to 38 µg/mL. This activity depends not only on the extract's chemical composition (analysed by HPLC/ESI-MS), but also on the target tumor cells. Interestingly, the cytotoxic effect of these extracts on the normal human peripheral blood mononuclear cells (PBMC) was weak when compared to that induced on tumor cells. On the other hand, oral route treatment of P815 tumor-bearing mice (DBA2/P815) with propolis ethanolic extract (5 mg per mouse every fourth day, five times for group A, and 2.5 mg per mouse every fourth day, five times for group B) significantly reduced the tumor volume (1.2 cm³ for group A and 2.7 cm³ for group B at the 22nd day after tumor graft). These effects are statistically significant as compared to those obtained with the control untreated mice (tumor volume 3.5 cm³ at day 22).

Ocular findings and histological change in glaucoma model of DBA/2J Mice / 眼科研究

Background As a hereditary chronic glaucoma model,DBA/2J mouse is widely used in the experimental research of glaucoma worldwide.Although some researches have determined the ocular change induced by hypertention of DBA/2J mouse,there is seldom reports about the research on the relationship of ocular pathological abnormality and development course in DBA/2J mouse.Objective The present study is to characterize the ocular abnormalities and histological changes induced by hypertention in different ages of DBA/2J mice.Methods The clean female DBA/2J mice aged 3-,5-,7-,9-,11-,14-month-old (6 mice for each) were used in this study,and age matched 18 female C57BL/6J mice were as controls.Intraocular pressure (IOP) of mice was measured by anterior chamber puncture of microneedle.The animals were sacrificed and retinal flat mounts were prepared for histopathological examination under the light microscope.Retinal ganglion cells (RGCs) were counted by retinal Nissl staining.Morphology of optic nerve cup in frozen section was examined under the light microscope.The experiment followed the Standard of Association for Research in Vision and Ophthalmology.Results Developing pigment dispersion,iris stroma atrophy,transillumination defects and pupil deformation were found in DBA/2J mice.IOP began to rise in 7-month-old DBA/2J mouse,peaked in 9-month-old mouse and returned to normal in 14-month-old DBA/2J mouse.A significant difference was found in IOP among different ages of DBA/2J mice (F=27.600,P＜0.05) but not C57BL/6J mice (F=0.249,P=0.781).RGCs loss was observed in 7-month-old DBA/2J mice and more serious from 9 to 11-month-old mice,showing a significant decline of RGCs among different ages of DBA/2J mice (F=23.594,P=0.000) but not C57BL/6J mice (F=1.816,P=0.211).The abnormality of optic nerve cupping and decrease of retinal nerve fiber layer were found in 9 to 14-month-old months old DBA/2J mice and were normal in age-matched C57BL/6J mice.Conclusion The abnormal alteration of the ocular anterior segment in DBA/2J mouse is gradually aggravated with aging.The findings of eye in DBA/2J mouse is characterized by secondary glaucoma.DBA/2J mouse offer an ideal animal model for the research of glaucoma.

Immunological regulation mechanism of epigallocatechin-3-gallate on collagen-induced arthritis / 中华风湿病学杂志

Objective The therapeutic effect of epigallocatechin-3-gallate (EGCG) on the collageninduced arthritis model (CIA) was observed and its immunological mechanism was analyzed. Methods EGCG was administered to CIA mice and PBS was admitted as negative control. The severity of CIA was evaluated by clinical scores and histopathological assessment (H-E staining). Immunological mechanisms inv-suppressive effect on IL-17 secretion of CD4+T cells (EGCG group: 0.41%; PBS group: 4.05% ) and inhibitive activity of C Ⅱ -reactive splenocytes proliferation. There was statistical significant difference between IKB expression and down-regulate phosphorylated IKB expression in lymph node cells of CIA mice.Conclusion EGCG can significantly ameliorate the severity of CIA. The therapeutic mechanisms may be related to inhibition of C Ⅱ -reactive splenocyte proliferation and IL-17 secretion and via inhibiting the activity of NF-κB by inducing the expression of IKB and by suppressing the expression of phosphorylated IKB in CIA mice.

Study on Protective Immunity against Infection of Plasmodium yoelii 17XL in DBA/2 Mice / 中国寄生虫学与寄生虫病杂志

Objective To investigate the development and dynamic changes of host immune response in DBA/2 mice infected with Plasmodium yoelii 17XL. Methods Female DBA/2 mice were infected by intraperitoneal ( i. p. ) injection of 106 P. yoelii 17XL parasitized erythrocytes ( PRBC). Levels of IL-12, IFN-γ, IL-4, IL-10 and P. yoelii 17XL-specific antibody in sera were measured by ELISA. Concentrations of NO in cell supernatants were measured by the Griess reaction. Parasitemia,percentage of mononuclear-macrophages of individual mice were monitored daily, and phagocytosis of mononuclear macrophages was also observed. Results Primary parasitemia in vein blood was developed on day 3 postinfection, which peaked with a level of 46. 9% on day 9. Most mice cleared the infection and survived by day 20 postinfection. From day 6 to day 16, the phagocytosis of PRBC by rodent macrophages was observed on the blood smear. Infected mice had a continuously increased level of IL-12 in serum from day 1 postinfection. Accordingly, high level of IFN-γ was also detected in sera from day 1 postinfection,which peaked on day 6. Infected mice produced higher level of IL-4 and IL-10 in serum on day 6 postinfection, which peaked on day 9 and day 15 postinfection respectively. In addition, splenocytes from infected mice produced significantly higher level of NO on day 6 and 20 postinfection. Level of P. yoelii 17XL-specific IgG was determined in the sera of infected mice with a steadily increased trend after infection, which peaked on day 70 postinfection. Conclusions Effective polarizing of Thl cells is significant in inhibition of parasitemia and eventual clearance of the Plasmodium parasites. Activated mononuclear-macrophages play a key role in inhibiting parasitemia in the early phase of infection with P. yoelii 17XL.

Effects of Triptolide on Bone and Joint of Mice with Collagen-lnduced Arthritis / 中国中医药信息杂志

Objective To observe the effect of triptolide on destruction of bone and joint of collageninduced arthritis (CIA) mouse. Methods Female DBA/1 mice were double immunized at the base of the tail with bovine type Ⅱ collagen (C Ⅱ). CIA mice were randomly divided into model group, methotrexate-treated group and 8.18, 16.36, 32.72 ?g/kg triptolide-treated groups. Performance of arthritis were observed regularly. Combined with radiologieal, histological methods were used, and the number of osteoelasts in bone was evaluated and analyzed with TRAP stainning. Results The clinical score and arthritis incidence of CIA mice were significantly decreased, and the number of osteoelasts in bone cavity reduced by treatment of triptolide. X-ray showed that the surface of bone of each articular was eroded, the joint space was narrow, BMD values of lumbar and knee were decreased compared with normal mice. Triptolide could significantly inhibit the damage of bones and joints of CIA mice, and increase the BMD values of lumbar and knee. Conclusion Triptolide can inhibit the damage of bones and joints of CIA mice.

Changes in Retinal Neuronal Population in DBA/2J Mice with Topical Intraocular Pressure Lowering Drugs

PURPOSE: The DBA/2J (D2) mouse is a transgenic mouse with pigmentary glaucoma. In a previous study, we found a reduction of inner retinal thickness in D2 mice. We attempted to discover the effect of eye drops on the retina of D2 mice. METHODS: Ten-month-old D2 mouse eyes were treated with Timoptic XE(R), Cosopt(R), and Xalacom(R) eye drops for a 1-month period. Immunohistochemical staining was performed on the mouse eye sections for analysis. RESULTS: In the control group, GABA and OPN immunoreactivity were markedly decreased and NOS immunoreactivity was increased. In all experimental group, GABA and OPN immunoreactivity were increased, and OPN immunoreactivity was markedly increased especially in the Cosopt(R) group. NOS immunoreactivity was decreased in all experimental groups. There was no difference in glycine immunoreactivity between the control and experimental groups. CONCLUSIONS: Combination anti-glaucoma eye-drops to the D2 mouse changed the retinal neuronal population and these drugs might play an important role in the mechanisms of retinal neuronal death; potential strategies for neuroprotection should therefore be evaluated.

Histologic Characteristics in the Eyes of DBA/2J Mice according to the Degree of Intraocular Pressure

PURPOSE: We assessed the correlation between ocular abnormalities and the degree of intraocular pressure (IOP) in the DBA/2J (D2) transgenic mice which were proven to have pigmentary dispersion syndrome and developing glaucoma. METHODS: Nine-months-old D2 mice were examined with biomicroscopy under anesthesia and measured for IOP by Tono-Pen, Hematoxylin and eosin staining was performed on the eye sections of the mice to analyze differences between the low-grade IOP group and the high-grade IOP group. RESULTS: Ocular abnormalities including iris pigment loss, iris transillumination, iris stromal atrophy, anterior synechia, thinning of the retina, and ganglion cell loss were found; all of which appeared to be pressure- dependent. CONCLUSIONS: These results corroborate that both IOP and age might be considered for studies using D2 mice, and suggest that D2 mice are a useful glaucoma model to study the mechanisms of retinal ganglion cell death and to evaluate strategies for neuroprotection.

Operating Rules and Regulations of SIEMENS dBA / 医疗卫生装备

With extensive application of interventional radiology technique, skilled and standardized manipulation of DSA is very helpful for interventional operations. It is introduced in this paper how to operate the Siemens dBA which is the first domestic one and settle with the problems met in routine work.

Histochemistry of Six Lectins in the Tissues of the Flat Fish Paralichthys olivaceus

Lectins are glycoproteins that specifically bind carbohydrate structures and may participate in the biodefense mechanisms of fish. In this study, the binding of three lectins, Dolichos biflorus agglutinin (DBA), soybean agglutinin (SBA), Bandeiraea simplicifolia BS-1 (isolectin B4), Triticum vulgaris (WGA), Arachis hypogaea (PNA) and Ulex europaeus (UEA-I) were studied in the gill, liver, intestine, kidney, heart, and spleen of the flat fish Paralichthys olivaceus. DBA was detected in intestinal mucous cells, as well as in gill epithelial and mucous cells. It was weakly detected in renal tubule epithelial cells and in bile duct epithelial cells. The strong SBA staining was seen in the intestinal club cells, in bile duct epithelial cells and renal tubule epithelial cells. There were intense positive reactions for isolectin B4 in gill epithelial and mucous cells, and the strong isolectin B4 staining was seen in epithelial cells of the bile duct and intestine. The strong WGA staining was seen in the gill mucosal cells, sinusoid, renal tubule epithelial cells and mucosal cells of the intestine. UEA-I was detected in the gill epithelial and mucosal cells, bile duct epithelial cells and renal tubular epithelial cells. These results suggest that the six lectins examined were localized in the covering epithelia of the various organs of the flat fish and they may participate in the biodefense mechanism of the intra body surface in which is exposed to various antigens.

Establishment and genetic checkup of an inbred strain of DXB/c mouse / 第三军医大学学报

An inbred strain of DXB/c mouse has been established by hybridization between DBA/2 female mice and C57BL/6 male ones and subsequently by sibmating their offsprings beginning from the F2 generation.Now DXB/c mouse has been passed for 28 generations of full sibmating since 1979.Genetic checkup by means of skin grafting,mandibular morphology analysis,mixed lymphocyte cultivation,coat colour gene testing,and biochemical marker gene examination confirmed that the full homozygosity of alleles has been achieved in DXB/c mouse and DXB/c mouse comforms to the criteria of an inbred strain of mouse.In addition,the genetic background of DXB/c mouse is composed of the genes of its progenitors DBA/2 and C57BL/6 as shown by coat colour gene testing and biochemical marker gene examination.

ANALYSIS OF THE G-BANDING PATTERN OF THE CHROMOSOMES OF THE DBA/2 MOUSE / 解剖学报

Bone marrow cells and cultures of embryo skin and lung cells from DBA/2 mice were obtained for chromosome analysis in our studies. The specimens were banded by Giemsa staining following trysinization, which produced well-scattered and sharply banded mitotic figures ranging from early metaphase to mid-metaphase. Over 435 bands Within the mouse karyotype can be distinguished. Idiogram of G-banding patterns were constructed on the basis of large amount of karyotype analysis. The features of banding patterns of the individual chromosomes are presented and those chromosomes with similar banding patterns are contrasted to avoid possible misidentification.