Role of Natural Killer Cell Subsets in Cardiac Allograft Rejection

To achieve donor-specific immune tolerance to allogeneic organ transplants, it is imperative to understand the cell types involved in acute allograft rejection. In wild-type mice, CD4(+) T cells are necessary and sufficient for acute rejection of cardiac allografts. However, when T-cell responses are suboptimal, such as in mice treated with costimulation-targeting agents or in CD28-deficient mice, and perhaps in transplanted patients taking immunosuppressive drugs, the participation of other lymphocytes such as CD8(+) T cells and NK1.1(+) cells becomes apparent. We found that host NK but not NKT cells were required for cardiac rejection. Ly49G2(+) NK cells suppressed rejection, whereas a subset of NK cells lacking inhibitory Ly49 receptors for donor MHC class I molecules was sufficient to promote rejection. Notably, rejection was independent of the activating receptors Ly49D and NKG2D. Finally, our experiments supported a mechanism by which NK cells promote expansion and effector function of alloreactive T cells. Thus, therapies aimed at specific subsets of NK cells may facilitate transplantation tolerance in settings of impaired T-cell function.     

Altered tachykinergic influence on gastric mechanical activity in mdx mice

This study investigated whether alterations in gastric activity in dystrophic mdx mouse can be attributed to dysfunctions of tachykinins. Endoluminal pressure was recorded and the expression of neuronal nitric oxide synthase (nNOS), NK1 and NK2 neurokinin receptors was investigated by immunohistochemistry. SR48968, NK2 receptor antagonist, but not SR140333, NK1 receptor antagonist, decreased the tone only in mdx gastric preparations. In the presence of N(omega)-nitro-l-arginine methyl ester (l-NAME), inhibitor of NOS, SR48968 reduced the tone also in normal stomach. [Sar(9), Met(O(2))(11)]-SP, agonist of NK1 receptors, caused tetrodotoxin-sensitive relaxations, antagonized by SR140333 or l-NAME, with no difference in the potency or efficacy between normal and mdx preparations. [beta-Ala(8)]-NKA(4-10), an NK2 receptor agonist, induced SR48968-sensitive contractions in both types of preparations, although the maximal response of mdx tissues was significantly lower than normal preparations. Immunohistochemistry demonstrated a consistent reduction of nNOS and NK2 receptor expression in mdx stomach smooth muscle cells and no change in nNOS and NK1 receptor expression in neurones. In conclusion, in mdx stomach the activation of NK2 receptors plays a role in the development of the tone, associated with a reduced NO production by muscular nNOS. The hypo-responsiveness to NK2 receptors could depend on the reduced expression of these receptors.     

抗端粒酶核酶抑制肝癌细胞裸鼠移植瘤生长的研究

设计针对端粒酶RNA组分的特异性核酶，研究该核酶对肝癌细胞裸鼠移植瘤生长的影响。构建含针对端粒酶RNA组分锤头状核酶基因的重组真核表达质粒pBBS212-RZ，以及建立人肝癌细胞株HepG2裸鼠移植瘤模型。将不同剂量的pBBS212-RZ用脂质体包裹后进行瘤体内多点注射，对照组注射生理盐水或空质粒载体pBBS212。连续注射2l天后，测量移植瘤体积和重量，检测瘤组织端粒酶活性。抗端粒酶特异性核酶使瘤组织端粒酶活性下降(抑制率为72％)，明显地抑制了肝癌细胞裸鼠移植瘤生长(抑制率为68％)，且存在剂量依赖性。抗端粒酶特异性核酶作为一种有效的端粒酶抑制剂，可抑制肝癌细胞的生长，有望在肝癌基因治疗中发挥作用。     

Isolation and differentiation of embryonic stem cells from BALB/c mouse

Objective To invest the efficient method which can culture and induce embryonic stem cells to neurocyte in vitro. Methods Isolate the blastula of 3.5 d from BALB/c species mouse. Culture the cells from inner cell mass (inner cell mass, ICM) which were isolated by mechanical method on the mouse embryonic fibroblaste cell (MEF) feeder layer or 0.1% gelatin coated dishes. The stem cells were identified by characterized morphology, alkaline phosphatase stain, differential potency in vivo and immunochemistry stain. The isolated cells were differentiated by serial induction method that mimicking the intrinsic developmental process of the neural system. Results The isolated cells were positive for alkaline phosphatatse and SSEA-1 (stage specific embryonic antigen 1). Moreover they were identified pluripotent by differentiation in vivo. Therefore the isolated cells presented the characters of ESCs. Then the isolated cells were able to differentiate into neurocytes in vitro. Conclusion Mouse embryonic stem cells isolation, culture and differentiation system has been established.     

Pollen extracts reduce the hepatotoxicity of paracetamol in mice

Cernitins are preparations obtained from plant pollen which contain numerous compounds of potential biological significance. This work deals with the influence of cernitins upon acute paracetamol toxicity in mice. The survival rate and indices of hepatic injury: aminotransferase and alkaline phosphatase activities, bilirubin level in serum, glutathione and cytochrome P-450 content in liver, liver weight, histopathologic picture of the liver and presence of glycogen and lipids in stained liver sections, under different experimental protocols, were determined. It was found that cernitins are able to increase the survival rate of mice and reduce liver injury in acute paracetamol poisoning. Cernitins are more effective when administered after, rather than before, a dose of paracetamol. The possible mechanism through which cernitins may act is discussed.     

Histochemical study of mitochondrial enzymes in cerebellar cortex of macular mutant mouse,a model of Menkes kinky hair disease

The cerebellar Purkinje cells in the hemizygote of the macular mutant mouse contain numerous abnormal mitochondria which show a marked decrease in cytochrome c oxidase activity. Using histochemical methods we studied the activity of other mitochondrial enzymes, such as NADH diaphorase and succinic dehydrogenase, in the cerebellar cortex of this mutant mouse. Such activities were markedly increased in the Purkinje cells, especially in the soma and stem dendrite, from 10 days after birth in the hemizygote as compared with findings in normal littermates. These results were considered to be due to an increased number of abnormal mitochondria.Supported by the Research Grants nos. 2A-5-03 and 3B-1-04 for Nervous and Mental Disorders from the Ministry of Health and Welfare, and by Grant no. 04670492 of the Ministry of Education of Japanese Government     

小鼠实验性糖尿病对胰腺腺泡细胞形态学的影响

为了探讨实验性糖尿病对胰腺腺泡细胞的影响，我们用四氧嘧啶对小鼠建造糖尿病模型。８ｗｋ后，取小鼠胰尾组织作光镜和电镜观察。结果：实验性糖尿病小鼠胰腺内部分腺泡细胞萎缩、细胞排列松散，并有大量慢性炎症细胞浸润。电镜下可见粗面内质网扩张，线粒体嵴紊乱、空泡变性，胞浆内出现髓样小体，酶原颗粒减少。表明实验性糖尿病对小鼠胰腺腺泡细胞的结构有一定的损害作用，进而使胰腺外分泌部的分泌功能受到影响。     

鸡矢藤提取物对小白鼠离体子宫的作用

鸡矢藤[Paederia Scandens]提取物0.32mg 加入营养槽内,对小白鼠各期离体子宫的收缩张力、收缩强度、收缩频率以及子宫活动力均有增强作用。与给药前比较,诱导期组分别增加85%、47%、21%和72%;动情期组分别增加70%、77%、41%和41%;非动情组分别增加47%、76%、62%和66%,而且三组间作用无显著性差异。本品还能增强 PGE_2对小白鼠离体子宫的收缩张力、收缩强度、收缩频率以及子宫活动力。与 PGE_2的作用比较,分别增加34%、39%、9%和45%,但收缩频率增加无统计学意义(P>0.05)。上述结果表明,鸡矢藤提取物对小鼠子宫有显著性兴奋作用,而且还能增强PGE_2对小鼠子宫兴奋作用,其机制尚待进一步研究。     

Experimental chronic dermatophytosis in human skin grafted to nude mouse: inoculation of Trichophytons and histopathological evaluation

Two strains of T. rubrum and one strain of T. mentagrophytes were inoculated into human skin grafted onto BALB/c nude mice by the needle puncture method. Infection was established in 1 of the 10 animals inoculated with fluffy colony type T. rubrum, 2 of the 10 animals inoculated with powdery colony type T. rubrum, and 7 of the 10 animals inoculated with granular colony type T. mentagrophytes, suggesting that the skin grafts are infectible by anthropophilic and zoophilic strains of dermatophytes. T. rubrum infection continued for a maximum of 9 weeks and T. mentagrophytes infection for more than 11 weeks. In the animals inoculated with T. mentagrophytes, fungal elements were localized in the stratum corneum of the human skin grafts. In the acute stage, microabscesses consisting of neutrophils were observed under the stratum corneum in contact with fungal elements; in the chronic stage, epidermal thickening and infiltration, mainly consisting of histiocytes and a smaller number of lymphocytes, was noted in the upper and middle dermis. Ultrastructural findings from the parasites were similar to those of dermatophytosis in man. This experimental system should be useful as a model of chronic dermatophyte infection in the human skin.