外源性p21WAF1/CIP1基因转染对人胶质瘤细胞生长和细胞周期的影响

目的：探讨外源性p21^WAF1／CIP1基因对胶质瘤细胞生长和细胞周期的影响．方法：采用脂质体介导法将p21^WAF1／CIP1基因质粒转导人人胶质瘤细胞系SHG44细胞，然后用电镜、流式细胞仪等技术对细胞形态、生长及细胞周期进行观察。结果：酶切电泳和斑点杂交显示p21^WAF1／CIP1基因成功的转染至SHG44细胞。转染p21^WAF1／CIP1基因的细胞光镜及电镜下可见凋亡产生；生长速度明显慢于亲代细胞，其倍增时间约是对照细胞的二倍；细胞周期发生明显改变，G1期明显增多，S期明显减少．结论：外源性p21^WAF1／CIP1基因对SHG44细胞的生长和细胞周期有明显的抑制作用，并可诱导细胞凋亡产生。     

Mental state,physical strength,intelligent cycle and their relationship with epilepsy

Patientsfeelimpatientbecausetheycouldnotpredicttheonsettimeofepilepsy．Ifonsettimeofepilepsycouldbepreciselypredicted，thendrugadministrationanddisabilityofpreventionwillbecomeavailable．In１８７９，Echevarrzafoundonsettimeofepilepsyisdistributedaccordingtosomerules．Inrecentyears，scientistsperformalotstudiesinvolvingcytologyandmolecularbiologyofepilepsy．Clinicaldataindicatedonsetofepilepsyhascloserelationwithbiologicalrhythmsuchasawakeandsleepcycleandmenstrualcycle．Theobjectiveofthisstudy…     

The estrous cycle affects cocaine self-administration on a progressive ratio schedule in rats

Although it has been demonstrated that many of the behavioral responses to psychomotor stimulants are gender dependent and hormonally sensitive, few studies have examined the possibility that the estrous cycle interacts with drug reinforcement in laboratory animals. The present experiment assessed the effect of the estrous cycle on two aspects of cocaine self-administration behavior: the breaking point on a progressive ratio (PR) schedule and the rate of cocaine intake on a fixed ratio one (FR1) schedule. On the PR schedule, the first lever response produced a drug infusion. Subsequent response requirements escalated with each injection until the behavior extinguished. Breaking points were defined as the final ratio completed. On a FR1 schedule, the estrous cycle had no effect on the rate of drug intake. On a PR schedule, female rats reached higher breaking points during estrus than during other stages of the estrous cycle. Furthermore, female rats displayed higher breaking points than male rats. It appears that the estrous cycle influences an animal's motivation to self-administer cocaine.     

RASSF1A基因表达对人肝癌细胞株QGY-7703生长的影响

目的观察RASSFlA基因的表达对人肝癌细胞株QGY-7703在体内外生长的影响。方法利用已构建成功的稳定表达RASSFlA基因的肝癌细胞株QGY-7703，通过细胞生长曲线、软琼脂集落形成试验、细胞周期的测定和裸鼠致瘤性试验对各稳定表达株的生物学行为进行检测。结果以未转染的QGY-7703为对照，RASSFlA基因的表达可明显抑制肝癌细胞在体外的生长速度，使肝癌细胞周期阻滞在G1／S期，显著降低肝癌细胞在软琼脂中形成的克隆数目（P〈0．01）和大小，显著减慢裸鼠皮下瘤的生长速度和重量（P〈0．01）。空载体的表达对肝癌细胞的生长影响不明显。结论RASSFlA的重表达抑制肝癌细胞在体内外的生长；RASSFlA基因是一个新的肝癌相关的抑癌基因。     

大鼠脊髓横断性损伤后细胞周期蛋白D3的表达变化

目的 探讨细胞周期蛋白D3（Cyclin D3）在横断性脊髓损伤（tSCI）后的表达变化以及定位情况。方法 将48只成年SD大鼠随机分为8组：正常对照组，T9横断伤2、8h、1、3、5、7和14d组，每组6只。采用Western blot测定损伤后各时间段Cyclin D3蛋白水平在脊髓中的表达变化。采用免疫组织化学方法检测Cyclin D3在正常以及损伤后脊髓中的分布和定位。结果 West—emblot显示，Cyclin D3蛋白水平在tSCI后头、尾段均呈现先升高后下降的趋势，尾段明显：Cyclin D3的表达于损伤后8h开始逐渐升高，3d达到高峰，一直持续到第5天，之后逐渐下降。免疫组织化学表明Cyclin D3在正常脊髓中均匀分布，损伤后3d，Cyclin D3在脊髓白质和灰质中表达明显增强；免疫荧光双标记表明Cyclin D3与神经元的标记物neuronal nucleus（NeuN）、少突胶质细胞标记物cyclic nucleotide 3’phosphohydrolase（CNPase）有明显共定位，与星形胶质细胞标记物gtial fibrillary acidic protein（GFAP）和小胶质细胞标记物OX-42也存在部分共定位。结论脊髓损伤后Cyc—lin D3蛋白水平呈现明显的时相变化，并且与神经元、少突胶质细胞、星形胶质细胞和小胶质细胞存在共定位，提示Cyclin D3参与了脊髓损伤后的病理生理过程。     

Hepatic UDP-glucose 13C isotopomers from [U-13C]glucose: a simple analysis by 13C NMR of urinary menthol glucuronide.

Menthol glucuronide was isolated from the urine of a healthy 70-kg female subject following ingestion of 400 mg of peppermint oil and 6 g of 99% [U-(13)C]glucose. Glucuronide (13)C-excess enrichment levels were 4-6% and thus provided high signal-to-noise ratios (SNRs) for confident assignment of (13)C-(13)C spin-coupled multiplet components within each (13)C resonance by (13)C NMR. The [U-(13)C]glucuronide isotopomer derived via direct pathway conversion of [U-(13)C]glucose to [U-(13)C]UDP-glucose was resolved from [1,2,3-(13)C(3)]- and [1,2-(13)C(2)]glucuronide isotopomers derived via Cori cycle or indirect pathway metabolism of [U-(13)C]glucose. In a second study, a group of four overnight-fasted patients (63 +/- 10 kg) with severe heart failure were given peppermint oil and infused with [U-(13)C]glucose for 4 hr (14 mg/kg prime, 0.12 mg/kg/min constant infusion) resulting in a steady-state plasma [U-(13)C]glucose enrichment of 4.6% +/- 0.6%. Menthol glucuronide was harvested and glucuronide (13)C-isotopomers were analyzed by (13)C NMR. [U-(13)C]glucuronide enrichment was 0.6% +/- 0.1%, and the sum of [1,2,3-(13)C(3)] and [1,2-(13)C(2)]glucuronide enrichments was 0.9% +/- 0.2%. From these data, flux of plasma glucose to hepatic UDPG was estimated to be 15% +/- 4% that of endogenous glucose production (EGP), and the Cori cycle accounted for at least 32% +/- 10% of GP.     

脑肿瘤干细胞体外分化过程中的回逆现象分析

目的探讨脑肿瘤干细胞（BTSCs）体外分化过程中的回逆现象，为研究其分化抑制机制奠定基础。方法利用CD133免疫磁珠筛选系统，从肿瘤组织中分离获得的CD133^＋细胞（BTSCs）分成4组进行培养：（1）含10％胎牛血清（FCS）；（2）10％FCS＋丙戊酸钠注射液（VPA）；（3）无FCS＋生长因子；（4）无FCS＋生长因子＋VPA。取不同时间点上的细胞，相差显微镜观察其形态变化：流式细胞术检测与分化相关的标志物、细胞周期和DNA倍体变化；利用免疫激光共聚焦分析与分化相关标志物的共表达情况。结果无FCS条件下培养的BTSCs呈悬浮球状生长，高表达CD133和巢蛋白（nestin），不表达胶质纤维酸性蛋白（GFAP）和β-微管蛋白Ⅲ（β-TubulinⅢ）。G0／G1期细胞占大多数，G2／M期细胞接近0％，DNA都是异倍体，对VPA反应不敏感。含FCS培养的原本悬浮的细胞约4h开始贴壁。均呈圆形。此后逐渐向多形性分化，至7d时分化的细胞部分又返回至圆形。至10d-21d时，有的还能重新恢复球形，并呈悬浮生长。培养3d、7d、10d和21d时，CD133、nestin阳性细胞数先降后升，GFAP^＋和β-TubulinⅢ^＋细胞数始终处于较低水平。含FCS培养液中加入VPA。细胞形态上未见上述的回逆现象，CD133和nestin表达的先降后升现象消失，GFAP和β-TubulinⅢ在第7天以后表达明显升高，但极大部分细胞共表达nestin。而神经干细胞（NSCs）在含FCS培养至10d时，即以GFAP和β-TubulinⅢ表达为主，未见CD133^＋细胞。此外，含血清培养时BTSCs仍以异倍体为主。含少量的G2／M期细胞，加VPA诱导后细胞周期和DNA倍体变化不明显。结论BTSCs在含血清条件下培养出现的多向分化表型不稳定，时有去分化所导致的回逆。加入诱导分化剂VPA培养，虽然能阻止回逆现象出现，并有代表星形胶质细胞和神经元标志物表达上升．但因其共表达nestin而仍属于未完全分化细胞，表明BTSCs分化始终处于受抑状态。     

Cutaneous postural vasoconstriction is modified by exogenous but not endogenous female hormones in young women

Previously reported attenuation of skin postural vasoconstriction in women during the luteal menstrual cycle phase may be due to a progesterone-mediated decrease in myogenic or veno-arteriolar (VAR) mechanisms. Skin perfusion was measured in the shin and foot dorsum by Laser Doppler Fluxometry during leg dependency that increased vascular transmural pressure below (myogenic constriction only) and above (myogenic and VAR) the 25 mmHg threshold for activation of the VAR, and during venous distension to activate the VAR alone (cuff inflation to 50 mmHg). In six young women with normal menstrual cycles, vasoconstrictor responses to all interventions did not differ between days 7–13 (follicular) and 18–23 (luteal) of the normal menstrual cycle when progesterone levels were low and at their peak respectively. In eight women taking combined oral contraceptives (OC) and tested during pill consumption days, reductions in foot skin perfusion were smaller (P = 0.05) than in the luteal phase of the normal cycle for leg dependency below (−36.9 ± 5.2% OC vs. −52.5 ± 7.8% luteal, mean ± S.E.M.) and above (−43.7 ± 3.4% OC vs. −55.1 ± 4.8% luteal) the VAR threshold, and for venous distension (−53.1 ± 2.6% OC vs. 66.4 ± 5.5% luteal). In women with normal menstrual cycles, impaired postural vasoconstriction may be confined to those who experience pre-menstrual symptoms rather than a direct effect of endogenous hormones. Reduced vasoconstriction in the dependent foot during OC use is consistent with the known vasodilator action of exogenous hormones and its long-term effects     

成年大鼠神经元和星形胶质细胞细胞周期蛋白依赖性激酶抑制因子表达的差异研究

目的比较研究成年大鼠细胞周期蛋白依赖性激酶抑制因子在神经元和星形胶质细胞的表达差异。方法应用免疫荧光和激光扫描共聚焦显微镜观察成年大鼠生理状态下大脑皮层或海马CA1、CA3、DG区神经元和星形胶质细胞细胞周期蛋白依赖性激酶抑制因子(CDKI)p15Ink4b、p21cipl的表达。结果成年大鼠海马区和大脑皮层的神经元有p15Ink4b和p21cipl的表达,细胞核和细胞浆均有表达,且以胞核为主;星形胶质细胞也有上述细胞周期调控蛋白的表达,便细胞数目较少,并且表达这些指标的星形胶质细胞多聚集在海马区。结论成年大鼠大脑皮层和海马区的神经元和星形胶质细胞均表达p15Ink4b和p21cipl,而其在神经元的表达较星形胶质细胞更为普遍。     

6186例脑卒中患者发病时间的季节性分析

目的探讨青岛市区脑出血(CH)、脑梗死(CI)、蛛网膜下腔出血(SAH)发病时间的季节规律。方法收集青岛市1998年～2002年脑卒中患者6186例,其中CH1179人,CI4821人,SAH186人。圆分布分析探讨脑卒中患者发病时间的季节规律。结果CH冬季高发,平均角在1月份(P<0.05),男性患者的平均角在2月份(P<0.05),女性患者的平均角在1月份(P<0.05);CI秋季高发,男、女间无差别,平均角在10月份(P<0.05);SAH秋季高发,平均角在10月份(P<0.05),男性患者的平均角在9月份(P<0.05),女性患者的平均角在10月份(P<0.05)。结论青岛市的脑卒中发病具有季节性,在脑卒中预防工作中要充分考虑其发病的季节特点,并采取相应的措施。