Effects of Reg‐2 on Survival of Spinal Cord Neurons In Vitro

Regeneration gene protein 2 (Reg‐2) is a small secreted protein expressed in motor and sensory neurons of spinal cord during developmental stages and following injury of peripheral nerves. Reg‐2 appears to act as a neurotrophic factor and protects injured neurons from death during regeneration. To illustrate these potential protective effects in vitro, we investigated the blocking effects of Reg‐2 antibodies on the survival of primary cultured spinal cord neurons and astrocytes, as well as on neurite outgrowth. In addition, the effects of Reg‐2 in neuron injury models induced by peroxide and mitochondrial poisoning were assessed. Our results showed that Reg‐2 antibody markedly reduced survival and neurite outgrowth from neurons, whereas astrocyte survival was unaffected. Addition of Reg‐2 into the culture medium had no effect on neuron survival or neurite outgrowth. However, the addition of the Reg‐2 into culture media after peroxide treatment or cellular hypoxia insult induced by mitochondrial poisoning can reduce lactate dehydrogenase release levels and cell death. Thus, the data suggests that Reg‐2 is essential for the survival and neurite outgrowth of developing spinal cord neurons but not the survival of glial cells, and that Reg‐2 plays protective effects on spinal cord neurons against injury in vitro. Anat Rec, 2010. © 2010 Wiley‐Liss, Inc.     

Development of new generation influenza vaccines : Recipes for success?

As infectious diseases cause approximately 25% of the annual global mortality, vaccines are found to be a time proven and promising response to infectious disease need. However, like for pharmaceutical small molecules, vaccine development is lengthy, risky and resource demanding. Faced with an attrition rate estimated around 80%, key opinion leaders were interviewed with the question: is there a recipe for success?     

血浆Reg3α蛋白水平与肠道aGVHD的相关性研究

本研究探讨血浆Re83d蛋白水平与异基因造血干细胞移植（allo—HSCT）后肠道急性移植物抗宿主病（GI—aGVHD）的临床诊断及预后的相关性。以我院2011年12月至2012年12月行allo—HSCT的103例患者为研究对象。采集移植前9d、0d、移植后14d、移植后28d、腹泻时多个时间点抗凝外周血,对发生aGVHD的患者进一步采集治疗1周及4周后抗凝外周血,采用ELISA方法检测血浆Reg3α蛋白浓度。结果表明,103例患者中无aGVHD17例,非aGVHD腹泻27例,单纯皮肤aGVHD10例,Ⅰ-Ⅱ度肠道aGVHD（GI—aGVHD）17例．Ⅲ-Ⅳ度GI—aGVHD32例。GI—aGVHD患者和非aGVHD腹泻患者腹泻时血浆Reg3et蛋白水平分别为111．5（54．7—180．2）ng／ml和23．9（14．5—89．5）ng／ml（P〈0．001）。Ⅲ-Ⅳ度GI—aGVHD患者治疗4周后,治疗有效组17例与治疗无效组7例血浆Reg3a蛋白水平分别为137．2（51．7—205．4）ng／ml和679．4（122．3—896．8）ng／ml（P=0．028）。治疗无效组7例全部死于aGVHD相关的多器官功能衰竭。血浆Reg3α蛋白浓度≥87．73ng／ml时预测Ⅲ-Ⅳ度GI—aGVHD的鉴别效度即曲线下面积最大（AUC=0．902）,其诊断Ⅲ-Ⅳ度GI—aGVHD敏感度为81．48％,特异性为82．86％。移植后Reg3α[蛋白高水平组（〈87．73ng／m1）患者Ⅲ-Ⅳ度GI—aGVHD发生率明显高于低水平组（〈87．73ng／m1）（P〈0．001）。结论：移植后血浆Reg3α蛋白水平升高提示Ⅲ-Ⅳ度GI—aGVHD的发生,Ⅲ-Ⅳ度GI—aGVHD患者免疫抑制治疗后血浆Reg3α蛋白水平不降低与预后不良相关,血浆Reg3α蛋白水平可作为提示GI—aGVHD的生物学标志物。     

Pancreatic regenerating protein (reg Ⅰ) and reg Ⅰ receptor mRNA are upregulated in rat pancreas after induction of acute pancreatitis

AIM: Pancreatic regenerating protein (reg Ⅰ) stimulates pancreatic regeneration after pancreatectomy and is mitogenic to ductal and β-cells. This suggests that reg Ⅰ and its receptor may play a role in recovery after pancreatic injury. We hypothesized that reg Ⅰ and its receptor are induced in acute pancreatitis.METHODS: Acute pancreatitis was induced in male Wistar rats by retrograde injection of 3% sodium taurocholate into the pancreatic duct. Pancreata and serum were collected 12, 24, and 36 hours after injection and from normal controls (4 rats/group). Reg Ⅰ receptor mRNA, serum reg Ⅰ protein, and tissue reg Ⅰ protein levels were determined by Northern analysis, enzymelinked immunosorbent assay (ELISA), and Western analysis, respectively. Immunohistochemistry was used to localize changes in reg Ⅰ and its receptor.RESULTS: Serum amylase levels and histology confirmed necrotizing pancreatitis in taurocholate treated rats. There was no statistically significant change in serum reg Ⅰ concentrations from controls. However,Western blot demonstrated increased tissue levels of reg Ⅰ at 24 and 36 h. This increase was localized primarily to the acinar cells and the ductal cells by immunohistochemistry. Northern blot demonstrated a significant increase in reg Ⅰ receptor mRNA expression with pancreatitis. Immunohistochemistry localized this increase to the ductal cells, islets, and acinar cells.CONCLUSION: Acute pancreatitis results in increased tissue reg Ⅰ protein levels localized to the acinar and ductal cells, and a parallel threefold induction of reg Ⅰ receptor in the ductal cells, islets, and acinar cells. These changes suggest that induction of reg Ⅰ and its receptor may be important for recovery from acute pancreatitis.     

Reg4在胃肠道肿瘤中的表达及意义

Reg4是再生基因家族（regenerating gene family）中的一员，定位于染色体1p12～13.1,表达一种由158个氨基酸组成的分泌性蛋白。Reg4蛋白主要表达于胃黏膜壁细胞和小肠上皮神经内分泌细胞，与胃肠道细胞的增殖分化有关。Reg4与人类胃肠道肿瘤的发生、演进、浸润、淋巴结转移、腹腔播散、5-氟脲嘧啶（5-FU）抗药性以及临床预后密切相关，为胃肠道肿瘤的临床诊断、用药指导及预后评估奠定了理论基础。     

HPLC法检测红景天贮存期红景天甙的含量

采用高效液相色谱法测定放置的狭叶红景天中红景天甙的含量。结果表明狭叶红景天存放一年内红景天甙含量没有变化。     

Serum olfactomedin 4 (GW112, hGC‐1) in combination with Reg IV is a highly sensitive biomarker for gastric cancer patients

Gastric cancer (GC) is 1 of the most common human cancers. Early detection remains the most promising approach to improving long‐term survival of patients with GC. We previously performed Serial Analysis of Gene Expression (SAGE) on 4 primary GCs and identified several GC‐specific genes including Reg IV. Of these genes, olfactomedin 4 (OLFM4, also known as GW112 or hGC‐1) is a candidate gene for cancer‐specific expression. In this study, we examined the expression of olfactomedin 4 in human GC by immunohistochemistry. We also assessed serum olfactomedin 4 levels in GC patients by enzyme‐linked immunosorbent assay. 94 (56%) of 167 GC cases were positive for olfactomedin 4 by immunostaining. Olfactomedin 4 staining was observed more frequently in stage I/II cases than in stage III/IV cases. The serum olfactomedin 4 concentration in presurgical GC patients (n = 123, mean ± SE, 36.3 ± 3.5 ng/mL) was significantly higher than that in healthy individuals (n = 76, 16.6 ± 1.6 ng/mL). In patients with stage I GC, the sensitivity of serum olfactomedin 4 (25%) and Reg IV (35%) was superior to that of CA19‐9 (5%) or CEA (3%). Furthermore, in patients with stage I GC, the combination of olfactomedin 4 and Reg IV elevated the diagnostic sensitivity to 52%. These results suggest that serum olfactomedin 4 is a useful marker for GC and its measurement alone or in combination with Reg IV has utility in the early detection of GC. © 2009 UICC     

RegⅣ、EGFR、PI3K蛋白在胃腺癌中的表达及意义

目的：研究再生基因蛋白Ⅳ（ Regenerating gene typeⅣ,RegⅣ）、表皮生长因子受体（ Epider-mal growth factor receptor ,EGFR）及磷脂酰肌醇3-激酶（ Phosphatidylinositol -3-kinase,PI3K）蛋白在胃腺癌中的表达及临床意义。方法应用S-P免疫组化技术检测73例胃腺癌及其相应的癌旁正常组织中RegⅣ、EGFR和PI3K蛋白的表达情况。结果73例胃腺癌组织中RegⅣ、EGFR、PI3K蛋白的阳性表达率分别为50．7％（37/73）、56．2％（41/73）、69．9％（51/73）均高于癌旁正常组织20．5％（15/73）、19．2％（14/73）、21．9％（16/73）,差异具有统计学意义（ P＜0．05）;RegⅣ的表达与肿瘤分化程度相关,差异有统计学意义（P＜0．05）;EGFR蛋白的表达与浸润深度、淋巴结转移、临床分期相关,差异有统计学意义（P＜0．05）;PI3K蛋白的表达与肿瘤分化程度、浸润深度、淋巴结转移、临床分期相关有统计学意义（P＜0．05）。 RegⅣ与EGFR、PI3K蛋白在胃腺癌组织中的表达呈正相关（r＝0．343、0．248,P＜0．05）,EGFR与PI3K蛋白在胃腺癌组织中的表达呈正相关（ r＝0．384,P＜0．05）。结论 RegⅣ可能通过激活EGFR/PI3K/Akt信号通路在胃腺癌的发生发展中发挥重要作用。     

胰岛β细胞参与急性胰腺炎后胰腺再生过程的研究

目的 探讨胰岛β细胞在实验性急性胰腺炎(AP)后胰腺再生过程中的作用.方法 SD大鼠87只,按数字表法随机分为对照组(15只)、糖尿病组(24只)、AP组(24只)和糖尿病+AP组(24只).采用腹腔注射链脲佐菌素(STZ,60 mg/kg体重)或左旋精氨酸(L-Arg,2.5 g/kg体重,2次)方法分别建立糖尿病和AP模型.术后1、3、5、7d分批处死大鼠.检测血淀粉酶和血糖水平;计算胰腺湿重比;胰腺组织常规病理学检查,计算胰腺坏死面积百分比和组织转化区域百分比;免疫荧光检测胰岛β细胞的再生基因( Reg4)和胰岛素的表达.结果 注射STZ后,大鼠血糖明显升高,注射L-Arg后大鼠胰腺组织水肿、坏死、炎细胞浸润,血淀粉酶明显升高,表明制模成功.制模后第3天糖尿病+ AP组的胰腺坏死面积为(71.6±6.0)％,显著大于AP组的(42.3±4.0)％;第7天的组织转化面积为(45.6±5.4)％,显著小于AP组的(78.5±6.4)％.糖尿病+AP组胰岛β细胞的Reg4和胰岛素表达均较AP组明显减少.结论 STZ破坏了胰岛β细胞,加重精氨酸诱导的AP的损伤,并抑制胰腺的再生过程.