高危妊娠与胎母输血的相关性分析

目的探讨孕晚期母亲血胎儿红细胞量（KB）与新生儿血红蛋白的相关性。方法运用用红细胞酸洗脱染色试验,监测297例入院待产孕妇的KB值,比较正常孕妇和高危孕妇KB的差异性,并观察其与新生儿出生后24 h内测的血红蛋白值的相关性。结果297例孕妇血中（正常妊娠162例,高危妊娠135例）,234例可测到胎儿红细胞,KB值为0.01%～5.2%,胎儿红细胞检出率占监测人数的78.79%。经非参检验表明,高危妊娠组的KB值明显高于正常组,差异有极显著意义（〈0.001）;母亲KB值与新生儿血色素值的相关性不强,但对母亲KB值进行分层观察发现,以KB≥0.4%为界值,≥0.4%与〈0.4%组间的新生儿Hb值存在差异显著性（=10.406,〈0.01）,KB≥0.4%组新生儿的Hb水平较低。结论大部分高危妊娠孕妇存在胎母输血现象。孕期KB值监测对胎母输血高危孕妇以及高危新生儿的筛选有一定价值。     

An increase of cytochrome C oxidase mediated disruption of gemcitabine incorporation into DNA in a resistant KB clone

Mechanistic aberrations leading to Gemcitabine (2',2'-dFdCyd,2,2-difluorodeoxycytidine, Gem) resistance may include alteration in its transport, metabolism and incorporation into DNA. To explore the mechanism of Gem resistance, the restriction fragment differential display PCR (RFDD-PCR) was employed to compare the mRNA expression patterns of KBGem (Gem resistant), KBHURs (hydroxyurea resistant) and KBwt (parental KB cell). Nine gene fragments were overexpressed specifically in the KBGem clone. Sequencing and BLAST results showed that three fragments represent cytochrome C oxidase (CCOX, respiration complex IV) subunit III (CCOX3). The cDNA microarray confirmed that the mRNAs of CCOX and ATP synthase subunits were upregulated in KBGem as compared to KBwt and KBHURs. The increase in CCOX1 protein and activity led to the increase of free ATP concentration, which is consistent with the gene expression profile of KBGem. Furthermore, the sensitivity to Gem could be reversed by sodium azide, a CCOX inhibitor. Following the treatment of sodium azide, the cellular accumulation of [3H]-Gem increased in a dose (of azide)-dependent manner, which is associated with increase of [3H]-Gem incorporation into DNA in KBGem. In summary, an increase of CCOX activity and free ATP level may reduce the transport, metabolism and DNA incorporation of Gem, resulting in Gem resistance.     

去卵巢2型糖尿病大鼠骨骼肌磷酯酰肌醇-3-激酶、丝氨酸／苏氨酸激酶2及核转录因子-kappa B的表达

目的探讨去卵巢2型糖尿病大鼠骨骼肌磷酯酰肌醇-3-激酶（PI3K）／丝氨酸／苏氨酸激酶2（Akt2）（PI3K／Akt2）信号通路相关因子及核转录因子-kappaB（NF—κB）表达的意义。方法29只雌性Wistar大鼠按随机数字表法分为对照组（NS,n=9）,2型糖尿病组（DS,n=10）,2型糖尿病去卵巢组（DOVX,n=10）,体重180～200g。NS组大鼠正常饮食,DS和DOVX组大鼠给予高糖高脂饮食8周后,予腹腔注射链脲佐菌素30mg／kg,糖尿病成模后1周,DOVX组大鼠摘除双侧卵巢,NS和DS组大鼠仅切除少量脂肪。去卯巢前后各组大鼠分别称重及测定空腹血糖、空腹胰岛素,计算胰岛素敏感指数。去卵巢12周后各组大鼠取两侧股四头肌分别行免疫组化及逆转录-多聚酶链反应（RT—PCR）检测PI3K、Akt2和NF—κB的表达水平。多组间比较采用方差分析,以P〈0．05为差异有统计学意义。结果（1）DS组大鼠骨骼肌PI3KmRNA和Akt2mRNA分别为0．797±0．043和0．753±0．076,低于NS组（0．870±0．051和0．844±0．013,t=2．722、3．368,均P〈0．05）,但高于DOVX组（0．715±0．068和0．666±0．038,t=2．686、2．526,均P〈0．05）。（2）DS组大鼠骨骼肌NF—κB mRNA为0．577±0．097,高于NS组（0．433±0．112,t=-3．147,P〈0．05）,但低于DOVX组（0．696±0．090,t=-2．862,P〈0．05）。（3）各组大鼠骨骼肌均可见P13K、Akt2和NF—κB蛋白表达。DS组大鼠骨骼肌Pi3K和Akt2蛋白分别为（4．3±0．6）和（3．4±0．7）,低于NS组（5．2±0．4、4．7±0．7,t=2．654、3．488,均P〈0．05）,但高于DOVX组（3．5±0．6和2．8±0．5,t=3．473、2．365,均p〈0．05）。（4）DS组大鼠骨骼肌NF—κB蛋白表达为6．5±0．9,高于NS组大鼠（4．9±0．8,t=-3．396,P〈0．05）,但低于DOVX组（8．1±0．8,t=-2．954,P〈0．05）。结论去卵巢2型糖尿病大鼠骨骼肌P13K和Akt2表达下降,NF—κB炎症因子激活可能导致胰岛素抵抗加重。     

核转录因子κB在大鼠全脑缺血耐受中的表达及意义

目的:探讨核转录因子кB(NFкB)在大鼠全脑缺血耐受中的表达及其意义。方法:建立大鼠四血管闭塞的全脑缺血耐受模型,将实验动物随机分为6组:缺血预处理组(IP),DTTC(NFкB的特异性抑制剂)加缺血预处理组(IP~(DTTC)),缺血再灌注组(I/R),缺血预处理加缺血再灌注组(IP+I/R),DTTC加缺血预处理组加缺血再灌注组(IP~(DTTC)+I/R),假手术组(sham)。各组行海马神经元计数,并用凝胶电泳迁移率改变分析法检测NFкB活性。结果:IP和I/R中NFкB均被激活;IP+I/R组与I/R组相比,神经元计数明显增高;DTTC可抑制NFкB活性,也抑制IP的神经保护作用。结论:IP可通过激活NFкB发挥神经保护作用。     

KB130015, A New Amiodarone Derivative with Multiple Effects on Cardiac Ion Channels

KB130015 (KB015), a new drug structurally related to amiodarone, has been proposed to have antiarrhythmic properties. In contrast to amiodarone, KB015 markedly slows the kinetics of inactivation of Na⁺ channels by enhancing concentration‐dependently (K_0.5# 2 μM) a slow‐inactivating I_Na component (#_slow# 50 ms) at the expense of the normal, fast‐inactivating component (#_fast# 2 to 3 ms). However, like amiodarone, KB015 slows the recovery from inactivation and causes a shift (K_0.5. # 6.9 μM) of the steady‐state voltage‐dependent inactivation to more negative potentials. Despite prolonging the opening of Na⁺ channels KB015 does not lengthen but often shortens the action potential duration (APD) in pig myocytes or in multicellular preparations. Only short APDs in mouse are markedly prolonged by KB015, which frequently induces early afterdepolarizations. KB015 has also an effect on other ion channels. It decreases the amplitude of the L‐type Ca²⁺ current (I^Ca‐L) without changing its time course, and it inhibits G‐protein gated and ATP‐gated K⁺ channels. Both the receptor‐activated I^k(ACH) (induced in atrial myocytes by either ACh, adenosine or sphingosylphosphorylcholine) and the receptor‐independent (GTPγS‐induced or background) I^k(ACH) are concentration‐dependently (K_0.5# 0.6 – 0.9 μM) inhibited by KB015. I^k(ATP), induced in atrial myocytes during metabolic inhibition with 2,4‐dinitrophenol (DNP), is equally suppressed. However, KB015 has no effect on I_K1 or on I_to. Consistent with the effects in K⁺ currents, KB015 does not depolarize the resting potential but antagonizes the APD shortening by muscarinic receptor activation or by DNP. Intracellular cell dialysis with KB015 has marginal or no effect on Na⁺ or K⁺ channels and does not prevent the effect of extracellularly applied drug, suggesting that KB015 interacts directly with channels at sites more easily accessible from the extracellular than the intracellular side of the membrane. At high concentrations KB015 exerts a positive inotropic action. It also interacts with thyroid hormone nuclear receptors. Its toxic effects remain largely unexplored, but it is well tolerated during chronic administration.     

Detection of bacteria-microtubule interactions in a cell-free extract

Host cell microtubules, or its primary constituent tubulin, participate in the cell-to-cell spread of some pathogenic bacteria. A cell-free method was devised for study of the interaction between these invasive bacteria and microtubules. The basis of the assay is a cytosolic extract of KB oral epithelial cells lysed by osmotic swelling and mechanical shearing. This technique is used in immunofluorscence micrography for visualization and an ELISA ideal for the screening of mutants deficient in the interaction.     

The enhanced radiation response of in vitro tumor models to insulin.

The presence of insulin during the irradiation of plateau phase baby hamster kidney, Chinese hamster ovary, Ehrlich tumor, human oral carcinoma, human laryngealcarcinoma, and V-79 cells improved radiation response. Insulin was also found to enhance the radiation response of multiceli V-79 spheroids. The radiosensitizing effect of insulin is both time and concentration dependent. Additional studies indicate that glucagon-free insulin and the insulin mimicking tripeptide, glyeyl-L-histidyl-L-lysine-acetate, also sensitize plateau phase cultures of V-79 cells to radiation.     

针刺督脉穴位对癫痫大鼠海马NF-κB表达的影响

目的探讨核因子-κB(NF-κB)在癫痫大鼠海马中的表达以及针刺督脉穴位治疗癫痫的机制。方法建立氯化锂-匹罗卡品癫痫大鼠模型,运用免疫组织化学方法观察大鼠海马中NF-κB的表达。结果空白组大鼠NF-κB免疫反应阳性细胞在细胞浆中分布较多,在细胞核内分布较少;模型组癫痫大鼠NF-κB免疫反应阳性细胞在细胞核中分布多;模型组癫痫大鼠与空白组大鼠的NF-κB比较差异有统计学意义(P<0.05);针刺组与模型组癫痫大鼠海马NF-κB比较差异有统计学意义(P<0.05)。结论癫痫发作后NF-κB被激活后转移至细胞核内,其表达明显增加。针刺督脉穴位可明显抑制癫痫大鼠海马NF-κB的表达,其机理可能是使刺激冲动传入中枢神经系统,适度抑制NF-κB表达,减轻其介导的炎症反应,抑制细胞过度凋亡,降低神经元兴奋性,达到抑制癫痫目的。     

COPD患者骨骼肌萎缩的分子生物学机制进展

慢性阻塞性肺疾病（COPD）不仅仅是一种局限于呼吸道和肺部的疾病,还是一种可以累及肺外各器官的全身疾病,特别是骨骼肌萎缩和功能障碍,严重影响患者的生活质量及预后。骨骼肌萎缩的机制尚不明确,其机制的发生是多种细胞因子及代谢通路共同参与的复杂过程。目前研究认为自噬、核因子KB、肿瘤坏死因子样弱凋亡诱导因子可能参与了COPD患者骨骼肌萎缩的发生,这些发现可能为COPD防治提供新的策略。     

妇科千金片对盆腔炎模型大鼠血清中TNF-α、NF-κB含量的影响

目的 探讨妇科千金片治疗盆腔炎的作用机理.方法 选用Wistar大鼠,运用子宫内接种混合菌液制造急性盆腔炎模型.随机分为空白组、假手术组、模型组、妇科千金片低、中、高剂量组、妇炎康组及罗红霉素组,治疗后,用LISA检测血清肿瘤坏死因子-α （TNF-α）、校因子κB （NF-kB）含量.结果 与空白组比较,模型组TNF-α、NF-κB的含量明显升高（P＜0.05）;与模型组比较,妇科千金片低、中、高剂量组、妇炎康组及罗红霉素组的TNF-α、NF-κB含量明显降低（P＜0.05）.结论 妇科千金片可能是通过调节血清中TNF-α、NF-κB的含量而发挥抗炎作用.