Inhibition of BDNF in Multiple Myeloma Blocks Osteoclastogenesis via Down-Regulated Stroma-Derived RANKL Expression Both In Vitro and In Vivo

Brain-derived neurotrophic factor (BDNF) was recently identified as a factor produced by multiple myeloma (MM) cells, which may contribute to bone resorption and disease progression in MM, though the molecular mechanism of this process is not well understood. The purpose of this study was to test the effect of BDNF on bone disease and growth of MM cells both in vitro and in vivo. Co- and triple-culture systems were implemented. The in vitro results demonstrate that BDNF augmented receptor activator of nuclear factor kappa B ligand (RANKL) expression in human bone marrow stromal cells, thus contributing to osteoclast formation. To further clarify the effect of BDNF on myeloma bone disease in vivo, ARH-77 cells were stably transfected with an antisense construct to BDNF (AS-ARH) or empty vector (EV-ARH) to test their capacity to induce MM bone disease in SCID–rab mice. Mice treated with AS-ARH cells were preserved, exhibited no radiologically identifiable lytic lesions and, unlike the controls treated with EV-ARH cells, lived longer and showed reduced tumor burden. Consistently, bones harboring AS-ARH cells showed marked reductions of RANKL expression and osteoclast density compared to the controls harboring EV-ARH cells. These results provide further support for the potential osteoclastogenic effects of BDNF, which may mediate stromal–MM cell interactions to upregulate RANKL secretion, in myeloma bone diseases.     

济南地区日光温室土壤养分的分布状况和累积规律

对济南地区不同种植年限日光温室内外及不同蔬菜作物的土壤养分差异和频数分布进行比较,分析了日光温室土壤养分随种植年限的累积特征,并对不同蔬菜温室土壤养分与种植年限进行曲线拟合,研究了引起土壤盐渍化和酸化的原因.结果表明:济南地区日光温室土壤碱解氮、速效磷、速效钾、有机质、电导率显著高于棚外土壤,增幅分别为135.3％、475.2％、290.1％、97.7％、188.7％,pH值较棚外土壤降低0.31;各土壤养分的频数分布曲线均呈正态分布.不同蔬菜温室土壤养分含量各有差别,其中,碱解氮和电导率为番茄＞甜椒＞黄瓜,有机质和pH为黄瓜＞甜椒＞番茄,速效磷为黄瓜＞番茄＞甜椒,速效钾为番茄＞黄瓜＞甜椒.日光温室土壤有酸化的趋势,但不明显.温室土壤盐渍化程度加重主要由碱解氮和速效钾的累积所致.pH的降低与碱解氮的累积关系密切.日光温室各土壤养分随种植年限的累积规律基本一致,1～2年为养分的快速累积期,3～4年的累积速率变缓,以后处于一个稳定水平,整体上表现为土壤系统的动态平衡.除pH的累积为负向外,其他均为正向累积.不同蔬菜温室土壤养分与种植年限呈极显著相关,且可用2次或3次曲线进行拟合.     

Long-term neuropeptide Y administration in the periphery induces abnormal baroreflex sensitivity and obesity in rats

Neuropeptide Y (NPY) is an important neuronal element involved in cardiovascular regulation. Since elevated plasma levels of NPY have been observed in numerous pathological situations, this study aimed to determine whether long-term elevated plasma concentrations of NPY could result in aberrant baroreflex sensitivity. Mini-osmotic pump containing NPY (85 μg per 30 days) was subcutaneously implanted between scapulae in male rats for 4 months. The rats treated with NPY showed the following characters compared with control group: (1) attenuated heart rate responding to the increases in mean arterial blood pressure (MABP) induced by phenylephrine, but enhanced heart rate responding to the decreases in MABP induced by sodium nitroprusside; (2) decreased protein levels of substance P (SP) and GluR2, while increased the expression of γ-aminobutyric acid A receptor (GABA(A)R) in brainstem; (3) abdominal obesity indicated by increased body weight and accumulated fat mass in peritoneal cavity; (4) significant increases in total cholesterol, triglycerides, and low density lipoprotein levels in the periphery. These findings indicate that long-term NPY administration in the periphery leads to abnormal baroreflex sensitivity due, at least in part, to the down-regulated expression of SP/GluR2 and elevated expression of GABA(A)R in both protein and RNA levels, which indicate the alternations in glutamate function and GABA action in the nucleus tractus solitarii in NPY-treated rats. Furthermore, long-term NPY administration results in abdominal obesity and dyslipidemia.     

Neuropeptide Y reverses chronic stress-induced baroreflex hypersensitivity in rats

Chronic stress, as a risk factor for cardiovascular diseases, has been reported to result in elevated plasma neuropeptide Y (NPY) and be highly associated with abnormal cardiac autonomic function. This study aimed to explore the effect of NPY on the chronic stress-induced abnormal baroreceptor reflex sensitivity (BRS). Seven types of recognized stressors were used to develop chronic stress rat model. Subcutaneously implanting ALZET mini-osmotic pumps containing NPY were used to evaluate the action of NPY on the stressed male rats. We found that chronic stress showed no influence on baseline systolic blood pressure (SBP) and heart rate (HR), whereas NPY (85 μg for 30 days) could elevate baseline SBP and induce bradycardia in rats intervened by various stimuli. NPY pretreatment could preserve chronic stress-induced decreases in left ventricular systolic pressure (LVSP) and the maximum rate of change in left ventricular pressure in the isovolumic contraction period (+dp/dt(max)) but has shown no effect on left ventricular end diastolic pressure (LVEDP) and the maximum rate of change in left ventricular pressure in the isovolumic relaxation period (-dp/dt(max)). Notably, chronic stress led to baroreflex oversensitivity indicated by the elevated ratio of Δheart rate (HR)/ Δmean arterial blood pressure (MABP) in rats followed by vasoconstrictor (phenylephrine, PE) or vasodilator (sodium nitroprusside, SNP) administration, which was almost completely reversed by NPY pretreatment. The expressions of substance P (SP) and gamma aminobutyric acid A receptor (GABA(A)R) in nucleus tractus solitarius were increased in chronic stress rats, which were counteracted by NPY pretreatment. We conclude that chronic stress-induced baroreflex hypersensitivity could be blocked by NPY pretreatment. Furthermore, the altered expressions of neurotransmitters and receptors in the brainstem might contribute to this process.     

半夏缓慢生长法保存及体细胞变异的ISSR检测

以添加了不同浓度的甘露醇、PP333和ABA的培养基对半夏试管苗进行缓慢生长法保存,并对保存材料再生后代的体细胞变异进行检测。结果显示,甘露醇、PP333和ABA均能有效抑制试管苗生长,且存活率高;最佳浓度分别为甘露醇2%～4%,PP3332.0mg·L-1,ABA 2.0～4.0mg·L-1。保存在添加了2%～4%甘露醇或2.0mg·L-1 PP333培养基上的植株未检测到变异,而保存在添加了2.0～4.0mg·L-1 ABA培养基上的植株检测到1条新增标记和1条缺失标记,位点变异率为1.7%,个体变异率为30%。研究表明,ABA不宜用于半夏试管苗的缓慢生长法保存,但有助于新突变体的产生,在种质创新上具有特殊意义。     

香石竹DcPAL1基因的克隆及其原核表达

苯丙氨酸解氨酶(PAL)是植物苯丙烷类代谢途径的关键酶。利用RT-PCR及RACE技术从香石竹茎中克隆到苯丙氨酸解氨酶基因的全长cDNA,命名为DcPAL1(GenBank登录号为FJ864719)。DcPAL1全长2 397bp,预测其开放阅读框长2 121bp,编码一个含有706个氨基酸的蛋白质,其分子量为76.8kD,等电点5.84,且含有PAL的特征序列和活性位点。构建pET-DcPAL1原核表达载体,并转入大肠杆菌(Escherichia coli)中表达,获得1个与预测大小一致的外源蛋白,主要以包涵体形式存在。用Ni2+-NTA螯合琼脂糖层析柱亲和层析得到了纯化的表达蛋白。     

雄性不育相关基因msLTP的反义RNA验证

根据普通白菜雄性不育相关的脂质转移蛋白基因(msLTP)的cDNA序列设计引物,从普通白菜花蕾的cDNA中扩增出312bp的片段,然后将该片段连接至双元载体pBI12l中,得到反义RNA植物表达载体并导入农杆菌LBA4404菌株中,通过农杆菌介导法转化菜心;利用PCR和Southern blot分析检测得到了25株转基因植株,转基因植株的花粉部分畸形或空瘪,花粉离体萌发率为38.56%,较未转化植株的萌发率(76.32%)降低了37.76个百分点.研究表明,反义RNA技术使msLTP基因沉默而导致了菜心转基因植株的部分花粉发育不良,说明msLTP基因在普通白菜和菜心等花粉发育中具有重要作用.     

The C-terminal α-helix of SPAS-1, a Caenorhabditis elegans spastin homologue, is crucial for microtubule severing

Spastin belongs to the meiotic subfamily, together with Vps4/SKD1, fidgetin and katanin, of AAA (ATPases associated with diverse cellular activities) proteins, and functions in microtubule severing. Interestingly, all members of this subgroup specifically contain an additional α-helix at the very C-terminal end. To understand the function of the C-terminal α-helix, we characterised its deletion mutants of SPAS-1, a Caenorhabditis elegans spastin homologue, in vitro and in vivo. We found that the C-terminal α-helix plays essential roles in ATP binding, ATP hydrolysing and microtubule severing activities. It is likely that the C-terminal α-helix is required for cellular functions of members of meiotic subgroup of AAA proteins, since the C-terminal α-helix of Vps4 is also important for assembly, ATPase activity and in vivo function mediated by ESCRT-III complexes.