Bcl-2和Caspase-9对儿童癫痫持续状态近期预后的预测价值

目的探讨血清B细胞淋巴瘤相关蛋白-2(Bcl-2)和半胱氨酸天冬氨酸蛋白酶-9(Caspase-9)对儿童癫痫持续状态(SE)近期预后的预测价值。 方法选择SE患儿62例为SE组,选择同期体检健康儿童62例为对照组;按照预后将SE患儿分为预后不良组和预后良好组。癫痫持续状态严重程度量表(STESS)评估SE患儿SE严重程度。ELISA检测各组血清Bcl-2和Caspase-9水平。结果SE组Bcl-2和Caspase-9水平高于对照组(P＜0.05)。预后不良组和预后良好组STESS评分、Bcl-2和Caspase-9水平比较,差异有显著性(P＜0.05)。STESS评分、Bcl-2和Caspase-9是影响SE患儿近期预后的独立影响因素(P＜0.05);入院血清Bcl-2和Caspase-9联合检测对SE患者近期预后具有较好的预测效能(P＜0.05)。 结论血清Bcl-2和Caspase-9是SE患儿近期预后的有效预测指标。     

SNHG6靶向miR-101/TGFBR1促进AMI小鼠左心室心肌纤维化

目的探讨SNHG6对急性心肌梗死(AMI)小鼠左心室心肌的影响。 方法将30只雄性C57/BL6小鼠构建成AMI小鼠后随机均分为AMI组、AMI+SNHG6组、AMI+miR-101-3p组、AMI+SNHG6+miR-101-3p组、AMI+miR-101-3p+TGFBR1组,另设正常小鼠6只为假手术组。qRT-PCR检测AMI小鼠SNHG6、miR-101-3p表达水平。心脏彩超检测各组小鼠左室射血分数(LVEF);马松和天狼星红染色法以及免疫组化分析各组小鼠左心室心肌纤维化变化。将H9C2细胞株分为阴性对照组(转染空质粒)、SNHG6组(转染质粒SNHG6)、miR-101-3p组(转染质粒miR-101-3p)。Western blotting检测各组TGFBR1蛋白表达;采用双荧光素酶报告基因法预测并验证SNHG6/miR-101-3p/TGFBR1荧光素酶活性及调控机制。 结果AMI小鼠较假手术组SNHG6表达显著增加,miR-101-3p降低(P＜0.05)。与AMI组比较,AMI+SNHG6组小鼠LVEF降低,心肌纤维化程度加重(P＜0.05);AMI+miR-101-3p组LVEF升高,心肌纤维化程度减轻(P＜0.05)。AMI+SNHG6+miR-101-3p组较AMI+SNHG6组LVEF升高、心肌纤维化程度减轻(P＜0.05),而AMI+miR-101-3p+TGFBR1组较AMI+miR-101-3p组LVEF降低、心肌纤维化程度加重(P＜0.05)。双荧光素酶报告基因法验证显示,miR-101-3p组SNHG6、TGFBR1野生型质粒的荧光素酶活性较阴性对照组明显降低(P＜0.05)。 结论SNHG6抑制miR-101-3p上调TGFBR1加重AMI小鼠左心室心肌纤维化。     

血管内皮细胞条件培养基促进胃癌细胞增殖和迁移

探讨血管内皮细胞培养基对胃癌细胞增殖和迁移的影响,分析血管内皮细胞调控胃癌细胞发生转移的机制。方法 设置对照组和共培养组,分别将正常培养基和人脐静脉血管内皮细胞HUVEC条件培养基作用于胃癌细胞HGC27,采用MTT法和划痕试验检测胃癌细胞HGC27的增殖活性和迁移能力。设置Control组、6 h组、12 h组和24 h组,以HUVEC的条件培养基作用于胃癌细胞6、12和24 h,Western blot检测EMT标志物和紧密连接蛋白的表达变化,激光共聚焦显微镜观察细胞骨架和紧密连接蛋白的分布变化。结果 MTT及划痕试验表明HUVEC条件培养基促进胃癌细胞HGC27的增殖和迁移。与Control组比较,间接共培养后胃癌细胞的形态呈间充质状改变,丝状伪足样凸起数量显著增加。 Western blot结果显示间接共培养后胃癌细胞上皮标志物E-cadherin表达水平逐渐下降,而间充质标志物N-cadherin和MMP-9逐渐增加,具有时序性变化规律。ZO-1和Occludin的表达也逐渐下降,细胞膜分布减少。结论 间接共培养下,血管内皮细胞通过上调胃癌细胞MMP-9,破坏紧密连接,促进胃癌细胞增殖和迁移     

Diffuse type of senile plaques in the cerebellum of Alzheimer-type dementia demonstrated byβ protein immunostain

Summary We studied senile plaques (SP) in the cerebella of six autopsied subjects with Alzheimer-type dementia (ATD) and ten non-ATD autopsied subjects between the ages of 78 and 90. Neither SP nor amyloid angiopathy (AA) was observed in any of the non-ATD subjects. In the four of the six ATD subjects, diffuse plaques in the molecular layer were seen as ill-defined areas of fine fibrillar materials by protein immunostaining with formic acid pretreatment, the modified Bielschowsky stain, and periodic acid-methenamine silver (PAM) stain. The plaques were not visible with Bodian, Congo red, or periodic acid-Schiff stains. Compact plaques in the Purkinje cell or in the granular cell layers were found in three of the six subjects. Their amyloid core was often surrounded by areolar amyloid deposits. AA was observed in three of the six subjects. The argyrophilia of the diffuse and compact plaques, demonstrated by the modified Bielschowsky and PAM stains, became undetectable when the sections were first treated with formic acid. Such treatment made the plaques immunoreactive with protein antiserum. The findings suggested that cerebellar diffuse plaques and compact plaques consist mainly of an amyloid component, and are characteristic of ATD.     

壬苯醇醚避孕药膜的临床研究

894对健康、已育的育龄夫妇,随机分两组:447对使用壬苯醇醚方型膜,447对使用帽型膜,经两年连续观察。按生命表统计法处理数据。总使用妇女月,方型组9601,帽型组9382,随访率100%。累计停用率、妊娠率和续用率,方型组分别为16.8%、15.3%和83.2%,帽型组为18.4%、15.6%和81.6%,两组无显著差异(p>0.05)。临床和实验室资料表明壬苯醇醚药膜是安全、有效、简便、易于推广的避孕药具之一。     

Characterization of NK cells and extrathymic T cells generated in the liver of irradiated mice with a liver shield

We previously reported that c-kit⁺ stem cells which give rise to extrathymic T cells are present in the liver of adult mice. Further characterization of extrathymic T cells in the liver of adult mice is conducted here. When mice with a liver shield were lethally (9.5 Gy) irradiated, all mice survived. All tested organs showed a distribution pattern of hepatic lymphocytes on day 7. The distribution pattern in the liver was characterized by an abundance of NK (CD3^? IL-2Rβ⁺) and extrathymic T cells (CD3^int IL-2Rβ⁺) before and after irradiation. To determine their function, post-irradiation allogeneic bone marrow transplantation (BMT) was performed in mice with or without a liver shield. Allogeneic BM cells were rejected in mice with a liver shield and specific activation of CD8⁺ CD3^int IL-2Rβ⁺ cells was induced. At that time, potent cytotoxicity of liver mononuclear cells (MNC) against allogeneic thymocytes was induced. Both NK1.1⁺ and NK1.1^? subsets of CD3^int cells expanded in these mice. An in vivo elimination experiment of the subsets indicated that the NK1.1⁺ subset of CD3^int cells (i.e. NK T cells) was much more associated with the rejection of allogeneic BM cells. However, even after the elimination of NK T cells, allogeneic BM cells were rejected. In this case, granulocytes expanded in parallel with NK1.1^? subsets. Granulocytes may also be associated with the rejection of allogeneic BM cells. These results suggest that the liver is an important haematopoietic organ even in adult life.     

Isolation and radiation hybrid mapping of dinucleotide repeat polymorphism at the human matrix Gla protein (MGP) locus

Matrix Gla protein (MGP) is an 84-residue, vitamin K-dependent protein expressed by chondrocytes and vascular smooth muscle cells, and is a potent regulator of calcium deposition in cartilage and arterial wall. We isolated a polymorphic dinucleotide CA repeat marker from a genomic clone containing the human MGP gene. This polymorphism will be useful in genetic studies of arteriosclerosis and osteoporosis. Received: November 5, 1997 / Accepted November 27, 1997     

Cannabinoid agonist, CP 55,940, facilitates intake of palatable foods when injected into the hindbrain

Cannabinoids have been shown to influence food intake, and until recently, the neural pathways mediating these effects have remained obscure. It has been previously shown that intracerebroventricular injection of delta-9-tetrahydrocannabinol (Δ⁹-THC) causes increased consumption of palatable foods in rats, and we postulated the involvement of the hindbrain in this cannabinoid-induced food intake. Cannulated rats (both female and male groups) trained to consume sweetened condensed milk received either lateral or fourth ventricle injections of CP 55,940 and were presented with sweetened condensed milk 15 min after injection. Rats were injected over a range of doses between 100 pg and 10 μg per rat. Milk intake was recorded for a total of 3 h. Lateral ventricle injection of CP 55,940 increased milk intake at doses in the microgram range. However, CP 55,940 was effective in increasing food intake at nanogram doses when injected into the fourth ventricle. Finally, male rats appeared to be more sensitive to CP 55,940 than female rats inasmuch as milk consumption was increased at the 1 ng dose in male rats, whereas only the 10 ng dose was effective in females. These results indicate that CP 55,940 may act in the hindbrain to influence feeding behavior in rats.     

Développement d’un hydrogel autodurcissant in vivo, en perspective d’un usage biomédicalA new self-hardening gel in prospect of biomedical use

Since 1995, an injectable bone substitute is developed in our laboratory, it is based on a mix of an hydrogel (hydroxypropyl methylcellulose at 3%: cellulose ether) and biphasic calcium phosphate granules (MBCP^® : 60% hydroxyapatite, 40% β-tri calcium phosphate). This first generation of injectable bone substitute is a non-self-hardening past, nevertheless it owns a great osteoconductor potential. But it shows a tendency to the discharge by its initial composition, and involved limited clinical applications. An evolution of this product is presented in this article: the modification is generated by a new self-hardening hydrogel. This hydrogel is a cellulose ether grafted by silane and diluted in an aqueous solution at basic pH. Also it will be presented the general synthesis of this cellulose derivate, the dissolution condition and the self hardening principle in function of the pH and the temperature. As to conclude by preliminary tests, the biocompatibility in vivo and in vitro of the self-hardening hydrogel mixed with biphasic calcium phosphate granules will be studied.     

Biosynthetic and regulatory role of lys9 mutants of Saccharomyces cerevisiae

Summary Derepression of lysine biosynthetic enzymes of Saccharomyces cerevisiae was investigated in lys9 auxotrophs which lack saccharopine reductase activity. Five enzymes (homocitrate synthase, homoisocitrate dehydrogenase, -aminoadipate aminotransferase, -aminoadipate reductase and saccharopine dehydrogenase) were constitutively derepressed in all lys9 mutants with up to eight-fold higher enzyme levels than in isogenic wild-type cells. Levels of these enzymes in lys2, lys14, and lys15 S mutants were the same or lower than those in wild-type cells. The regulatory property of lys9 mutants exhibited recessiveness to the wild-type gene in heterozygous diploids. Unlike the mating type effect, homozygous diploids resulting from crosses between lys9 auxotrophs exhibited even higher levels of derepressed enzymes than the haploid mutants. Addition of a higher concentration of lysine to the growth medium resulted in reduction of enzyme levels although they were still derepressed. These results suggest that lys9 mutants represent a lesion for the saccharopine reductase and may represent a repressor mutation which in the wild-type cells simultaneously represses unlinked structural genes that encode for five of the lysine biosynthetic enzymes.