泽兰有效部分对血小板聚集和血栓形成的影响

目的 研究泽兰有效部分L．F04：对血小板聚集和血栓形成的影响，探讨其活血化瘀作用机制。方法用高分子右旋糖酐静脉推注造成大鼠血瘀证模型，观察泽兰L.F04对二磷酸腺苷(ADP)诱导的大鼠体内血小板聚集以及体内动静脉旁路血栓、体外旋转环内血栓形成的影响。结果 L．F04 0．408，0．204g／kg对模型组大鼠ADP诱导的体内血小板最大聚集率明显增加皆有显著的抑制。且呈剂量依赖关系；与对照组相比，血瘀模型大鼠体外血栓质量明显增加，长度仅有增加趋势，L．F04高、低剂量(0．408，0．204g／kg)皆有抗血栓形成作用，L．F04高剂量对血栓干质量、湿质量的减轻尤为明显；L．F04高、低剂量对实验性动静脉旁路血栓形成均有明显的抑制作用，抑制率分别为27．41％，27．14％。结论泽兰L．F04可显著抑制血小板聚集及体内、外血栓形成。     

Coordinated age-dependent and pancreatic-specific expression of mouse Reg2Reg3α, and Reg3β genes

Reg family proteins such as Reg1 and islet neogenesis-associated protein (INGAP) have long been implicated in the growth and/or neogenesis of pancreatic islet cells. Recent reports further suggest similar roles to be played by new members such as Reg2, Reg3α, and Reg3β. We have studied their age-, isoform-, and tissue-specific expressions. RNA and protein were isolated from C57BL/6 mice aged 7, 30, and 90 days. Using real-time polymerase chain reaction, the levels of Reg gene expression in the pancreas were 20–600-fold higher than that in other tissues (?duodenum>stomach>liver); gene expression of Reg2, Reg3α, and Reg3β was age dependent as it was hardly detectable at day 7, increased drastically at day 30, and significantly decreased at day 90; the levels of pancreatic proteins displayed similar age-dependent variations. Using dual-labeled immunofluorescence, Reg2, Reg3α, and Reg3β were abundantly expressed in most acinar cells of the pancreas, in contrast to INGAP which exhibited stepwise increases from day 7 to day 90 and colocalized with the α-cells. These new Reg genes were mainly expressed in the pancreas, with clear age-dependent and isoform-specific patterns.     

Expression of reg protein in rat regenerating islets and its co-localization with insulin in the Beta cell secretory granules

Summary Regenerating islets can be induced by the administration of poly(ADP-ribose) synthetase inhibitors to 90% depancreatized rats. In screening a regenerating isletderived cDNA library, we previously isolated a novel gene, reg (regenerating gene), which encodes a 165-amino acid protein with a 21-amino acid signal sequence. In the present study, we have examined the expression and localization of reg protein in the regenerating islets by immunocytochemical techniques using a monoclonal antibody against a recombinant rat reg protein of 144 amino acids without the signal sequence. Light microscopy examination showed strong immunoreactivity for reg protein in the regenerating islets of the rats at two weeks and two months after the 90% pancreatectomy, whereas reg protein was almost undetectable in normal rat islets or in the islets of the rats one year after the pancreatectomy. Almost all the reg protein-positive cells were stained for insulin. By applying the immunogold technique at the ultrastructural level, it was demonstrated that both reg protein and insulin occur in the central granular core of the regenerating Beta cell secretory granules. These results suggest that reg protein is synthesized in and secreted from the regenerating Beta cells and that its expression is closely associated with Beta-cell regeneration.     

The inhibitory principle of lipopolysaccharide-induced nitric oxide production from<Emphasis Type="Italic">inula britannica</Emphasis> var.<Emphasis Type="Italic">chinensis</Emphasis>

A sesquiterpene lactone, 1-O-acetyl-4R,6S-britannilactone (1) isolated from the flowers of Inula britannica L. var. chinensis (Rupr.) Reg. (Compositae), was found as an iNOS inhibitory constituent for the first time with an IC50 value of 22.1 microM which is more potent than the positive control, L-N6-(1-iminoethyl)lysine (IC50 = 33.7 microM). Structure of compound 1 was identified by 1D and 2D NMR experiments and by comparison with the reference standard.     

Reg IV,a new member of the regenerating gene family,is overexpressed in colorectal carcinomas

A better understanding of the mechanisms by which colon tumor cells are able to survive exposure to drugs would be valuable for the development of new therapeutic strategies. We used differential display-PCR to compare gene expression in the drug-sensitive HT-29 colon cancer cell line and 3 drug-resistant subpopulations derived from this parental cell line. One of the genes identified is a new gene, Regenerating IV gene (Reg IV), and was strongly overexpressed in HT-29 drug-resistant cells. Other drug-resistant cell lines expressed Reg IV at a high level, whereas a low expression was noted in sensitive cell lines. Northern blot and real-time PCR analysis showed that Reg IV is more strongly expressed in 71% of colorectal tumors (in particular in mucinous carcinomas) than in normal colon tissues. The comparison of Reg IV expression with that of other REG genes, Regenerating Ialpha or (Reg Ialpha), Regenerating Ibeta (Reg Ibeta) and Pancreatitis-associated protein (PAP), highlights its predominant expression in colorectal tumors. Reg IV mRNA-positive tumor cells display different phenotypes: mucus-secreting, enterocyte-like or undifferentiated. Interestingly, whereas Reg IV expression is low in normal colon, its level in normal small intestine is similar to that in some colorectal tumors. In normal tissue, Reg IV mRNA-positive cells are mostly enteroendocrine cells and goblet cells. Our results point out the potential role of Reg IV in colorectal tumors and its subsequent interest as a pronostic indicator of tumor survival.     

Reg IV expression is associated with cell growth and prognosis of adenoid cystic carcinoma in the salivary gland

Aims: Regenerating islet‐derived family, member 4 (Reg IV) is associated with the progression of various cancers. The aim was to examine Reg IV expression in adenoid cystic carcinomas (ACCs) in salivary glands. Methods and results: Reg IV expression was detected by immunohistochemistry and compared with clinicopathological parameters. Expression of phosphorylated epidermal growth factor receptor (pEGFR), phosphorylated AKT (pAKT) and MUC2 was examined by immunohistochemistry. Reg IV function was assessed with Reg IV antisense S‐oligodeoxynucleotides (AS) in ACC3 human ACC cells. Reg IV was expressed by salivary duct epithelia and acinus myoepithelia, but not in squamous epithelia. Reg IV expression was found in 41% (17/41) of ACCs, but in none of 40 oral squamous cell carcinomas (OSCCs) and was associated with nodal metastasis (P = 0.047) and poor prognosis (P = 0.012) in ACCs. Reg IV expression was associated with pEGFR (14/17, 82%) in Reg IV + ACCs, but had no relationship with pAKT or MUC2 expression in ACCs. Cell growth was inhibited by AS treatment in Reg IV + ACC3 cells, but not in HSC‐4 OSCC cell s , whereas in vitro invasion of neither cell types was affected by AS treatment. Conclusions: These results suggest that Reg IV might accelerate cell growth and disease progression of ACCs.     

IL-21 signaling is critical for the development of type I diabetes in the NOD mouse

IL-21 is a pleiotropic type I cytokine that shares the common cytokine receptor γ chain and plays important roles for normal Ig production, terminal B cell differentiation to plasma cells, and Th17 differentiation. IL-21 is elevated in several autoimmune diseases, and blocking its action has attenuated disease in MRL/lpr mice and in collagen-induced arthritis. The diabetes-associated Idd3 locus is at the Il2/Il21 locus, and elevated IL-21 was observed in the nonobese diabetic (NOD) mouse and suggested to contribute to diabetes by augmenting T cell homeostatic proliferation. To determine the role of IL-21 in diabetes, Il21r-knockout (KO) mice were backcrossed to NOD mice. These mice were devoid of lymphocytic infiltration into the pancreas, and only 1 of 20 animals had an elevated glucose compared with 60% of NOD mice on a wild-type (WT) background. Although TCR and Treg-related responses were normal, these mice had reduced Th17 cells and significantly higher levels of mRNAs encoding members of the Reg (regenerating) gene family whose transgenic expression protects against diabetes. Our studies establish a critical role for IL-21 in the development of type I diabetes in the NOD mouse, with obvious potential implications for type I diabetes in humans.     

PBudCE4．1／Reg基因联合胰岛素自身抗原防治1型糖尿病的作用研究

目的探讨PBudCE4．1／Reg基因联合胰岛素自身抗原共同防治1型糖尿病的作用及机制。方法构建PBudCE4．1／Reg基因;采用40mg／kg链脲佐菌素（STZ）连续5d腹腔注射建立1型糖尿病小鼠模型,小鼠每周肌肉注射Reg基因,皮下注射胰岛素不完全弗氏佐剂（IFA）混合乳剂各1次,连续4周。每周测定小鼠血糖,6周后处死小鼠。采用四唑盐比色法（MTT）检测小鼠脾淋巴细胞增殖反应;采用流式细胞术（FACS）分析脾淋巴细胞CD4^＋CD25^＋Foxp3^＋调节性T细胞阳性率;采用ELISA法检测小鼠血清胰岛素分泌水平;采用HE染色法进行胰腺组织病理组织学检查。结果Reg／胰岛素联合给药可有效减少胰岛细胞损伤及炎性细胞浸润,提高糖尿病小鼠胰岛素分泌水平,从而显著降低糖尿病小鼠血糖值（P〈O．01）,且停药2周后,降糖作用仍持续存在;同时可降低脾淋巴细胞增殖能力（P〈0．01）,促进CD4^＋CD25^＋Foxp3^＋调节性T细胞分化（P〈0．05）。结论Reg基因联合胰岛素自身抗原对STZ诱导的T1DM模型有明显的抗糖尿病作用及协同效应,作用机制可能与免疫耐受的诱导及B细胞修复再生有关。