康莱特对Patu-8988细胞周期及其调节基因表达的影响

目的研究康莱特注射液对人胰腺癌细胞周期及其调节基因表达的影响,探讨康莱特的药理机制.方法通过流式细胞仪DNA含量分析法检测康莱特对人胰腺癌细胞系Patu-8988细胞周期的影响,应用基因芯片技术分析加药前后细胞周期调节基因的表达差异,并以Western blot对部分基因蛋白产物的表达进行验证.结果 20 μl/ml康莱特作用Patu-8988细胞24 h后,G2/M期细胞比率从(17.79 ± 0.16)%上升至(23.96 ± 2.33)%,而S期细胞比率则由(36.61 ± 2.97)%降至(24.76 ± 4.92)%.基因芯片结果显示,在96条有关细胞周期的目的基因中共有27条基因表达发生大于3倍的显著变化,其中表达上调的基因17条,下调的10条.Western blot表明cyclin A、cyclin B1、cyclin E、p21等蛋白表达改变与基因芯片结果一致.结论康莱特的药理作用之一是使细胞周期相关基因的表达发生改变,从而影响细胞增殖.     

康莱特对Patu-8988细胞周期及其调节基因表达的影响

目的　研究康莱特注射液对人胰腺癌细胞周期及其调节基因表达的影响 ,探讨康莱特的药理机制。方法　通过流式细胞仪 DNA含量分析法检测康莱特对人胰腺癌细胞系 Patu- 8988细胞周期的影响 ,应用基因芯片技术分析加药前后细胞周期调节基因的表达差异 ,并以 Western blot对部分基因蛋白产物的表达进行验证。结果　 2 0 μl/ ml康莱特作用 Patu- 8988细胞 2 4 h后 ,G2 / M期细胞比率从(17.79± 0 .16 ) %上升至 (2 3.96 ± 2 .33) % ,而 S期细胞比率则由 (36 .6 1± 2 .97) %降至 (2 4 .76 ±4 .92 ) %。基因芯片结果显示 ,在 96条有关细胞周期的目的基因中共有 2 7条基因表达发生大于 3倍的显著变化 ,其中表达上调的基因 17条 ,下调的 10条。Western blot表明 cyclin A、cyclin B1、cyclin E、p2 1等蛋白表达改变与基因芯片结果一致。结论　康莱特的药理作用之一是使细胞周期相关基因的表达发生改变 ,从而影响细胞增殖。     

SIRT1抑制剂sirtinol对前列腺癌DU145细胞凋亡的影响及可能机制

目的观察SIRT1抑制剂sirtinol对前列腺癌DU145细胞系细胞凋亡和细胞凋亡关键调控因子(Bcl-2、Bax、Cytochrome C和Caspase-3)表达变化的影响,探讨SIRT1在前列腺癌发生的可能机制。方法体外培养DU145细胞,实验分对照组(DMSO组)和sirtinol组(终浓度为10,25和50μmol/L),Western印迹检测DU145细胞SIRT1蛋白表达水平;流式细胞术检测细胞凋亡;Western印迹检测DU145细胞中细胞凋亡关键调控因子Bcl-2、Bax、Cytochrome C和Caspase-3的蛋白表达。结果与对照组相比,随着sirtinol浓度的增加,SIRT1蛋白表达水平逐渐降低,细胞凋亡比例增加,诱导细胞发生凋亡。不同浓度sirtinol作用DU145细胞后Bcl-2蛋白表达减少,且随剂量增加表达越低;Bax蛋白表达增加,Bcl-2/Bax比率降低;伴随Cytochrome C的释放和Caspase-3的激活。结论下调SIRT1表达诱导前列腺癌细胞DU145细胞凋亡,其机制可能与改变细胞凋亡关键调控因子Bcl-2、Bax、Cytochrome C和Caspase-3的蛋白表达相关。     

沉默Survivin人耐药肺癌细胞凋亡相关基因表达变化及临床意义

目的探讨沉默Survivin基因后人耐药肺腺癌(A549DDP)细胞系凋亡相关基因表达变化及临床意义。方法应用RT-PCR法及Western印迹法比较Survivin在人肺腺癌A549细胞系及其耐药A549DDP细胞系的表达水平;通过小分子RNA干扰沉默Survivin基因,并采用微阵列PCR基因芯片(PCR-Array)技术比较沉默Survivin后A549DDP细胞凋亡相关基因表达水平变化。结果 (1)Survivin mRNA、Survivin蛋白在人耐药非小细胞肺癌细胞系呈现高表达。(2)沉默Survivin后A549DDP细胞部分抗/促凋亡基因表达发生改变,促凋亡基因TP53、CASP3、CASP7呈高表达,抗凋亡基因bcl-2、TRAF1、BFAR呈低表达。结论 (1)Survivin与部分抗/促凋亡基因共同参与了非小细胞肺癌耐药的发生。(2)封闭Survivin可改变耐药肺癌细胞的促/抗凋亡基因的表达,Survivin可能作为上游调控点参与调控部分促/抗凋亡基因组的表达。(3)Survivin可能成为逆转肺癌耐药的一个新靶点。     

孕烷X受体抗食管癌EC9706细胞凋亡的作用机制

目的 研究孕烷X受体(PXR)抗食管癌EC9706细胞凋亡的作用机制.方法 使用利福平活化食管鳞癌EC9706细胞中的PXR,阿霉素(ADM)诱导高表达PXR的EC9706细胞凋亡,采用流式细胞仪观察细胞的增殖周期,MTT法观察细胞凋亡率,Western印迹和免疫组化法检测Caspase-3,Bcl-2,Bax蛋白表达情况.结果 ADM处理可以明显抑制细胞生长;使细胞呈明显凋亡改变;利福平诱导PXR高表达的EC9706细胞凋亡减少,抑制Caspase-3的蛋白水平,上调蛋白Bcl-2的表达,表明PXR在抗食管鳞癌细胞凋亡中发挥重要的作用.结论 PXR可能是通过降低Caspase-3和升高Bcl-2蛋白的表达抑制食管癌细胞EC9706的凋亡.     

芹菜素对膀胱癌T24细胞增殖、凋亡和CyclinD1、Caspase-3表达的影响

目的研究芹菜素(apigenin)诱导膀胱癌T24细胞凋亡的作用,探讨其作用机制。方法 CCK8法测定不同浓度芹菜素对T24细胞生长情况的影响;流式细胞术检测T24细胞周期情况;AnnexinV-FITC/PI双染流式细胞术观察T24细胞凋亡情况;Western印迹法检测CyclinD1和Caspase-3蛋白表达的变化。结果芹菜素能诱导T24细胞凋亡(P<0.05),并明显抑制CyclinD1的表达(P<0.05,P<0.01),同时上调Caspase-3的表达(P<0.05,P<0.01)。结论芹菜素在体外可抑制T24细胞增殖,诱导其发生凋亡。其作用机制可能与CyclinD1的表达下降,Caspase-3的表达增加有关。     

顺铂对人舌鳞状细胞癌细胞Tca8113细胞凋亡与自噬的影响

目的 探讨顺铂对人舌鳞状细胞癌细胞Tca8113细胞凋亡与自噬的影响.方法 体外培养人舌鳞状细胞癌Tca8113细胞,给予不同浓度的顺铂(CDDP),通过MTT检测细胞增殖率;Western印迹检测CDDP对Tca8113细胞凋亡以及自噬相关蛋白表达水平的影响.结果 与对照组相比,CDDP可以明显降低Tca8113细胞增殖率(P＜0.05);Western印迹结果显示,CDDP可以明显增加凋亡相关蛋白Cleaved Caspase-3,Cleaved PARP以及Cytochrome C的表达水平;同时,自噬相关蛋白LC3,Beclin 1和p62表达水平也发生明显的自噬性改变.结论 CDDP可以明显抑制Tca8113细胞增殖,其增殖抑制作用可能来源于凋亡,同时CDDP可以诱导Tca8113细胞发生自噬.     

蓝莓花色苷对人脐静脉内皮细胞凋亡调控基因Bax、Caspase-3表达的影响

目的探讨蓝莓花色苷对血管紧张素(Ang)Ⅱ诱导的人脐静脉内皮细胞(HUVEC)凋亡及凋亡相关基因表达的影响。方法将HUVEC常规复苏传代,建立HUVEC凋亡模型,随机分成空白对照组、AngⅡ组、蓝莓花色苷组、AngⅡ+蓝莓花色苷组。采用MTT法检测细胞活性,通过Annexin V-FITC标记的流式细胞术检测细胞凋亡率,Western印迹蛋白免疫印迹法检测Bax、Caspase-3蛋白表达。结果与空白对照组相比,AngⅡ组Bax和Caspase-3蛋白表达水平显著增加(P0.01);与AngⅡ组相比,蓝莓花色苷组、AngⅡ+蓝莓花色苷组Bax和Caspase-3的蛋白表达水平明显降低(P0.01)。结论蓝莓花色苷对HUVEC凋亡有一定抑制作用。     

X盒结合蛋白1在高迁移率族蛋白B1

目的　探讨活化的X盒结合蛋白1（XBP1）在高迁移率族蛋白B1（HMGB1）诱导人脐静脉血管内皮细胞凋亡中的作用及机制。方法　体外培养人脐静脉血管内皮细胞,将其分为正常对照组、HMGB1刺激组和慢病毒干扰＋HMGB1刺激组（即XBP1基因被沉默后加入HMGB1）。采用RT-PCR检测剪切型XBP1（sXBP1）的基因表达水平,Western　blot检测Caspase-3的蛋白表达水平,细胞荧光染色检测内皮细胞凋亡。结果　与正常对照组比较,HMGB1刺激组sXBP1的基因表达水平增加,Caspase-3的蛋白表达水平明显升高,细胞凋亡率明显增加（P均＜0.05）;与正常对照组比较,慢病毒干扰＋HMGB1刺激组sXBP1的基因表达水平无明显改变,Caspase-3的蛋白表达水平、细胞凋亡率差异无统计学意义（P>0.05）。与HMGB1刺激组比较,慢病毒干扰＋HMGB1刺激组sXBP1的基因表达水平降低,Caspase-3的蛋白表达水平降低,细胞凋亡率降低（P均＜0.05）。结论　XBP1对HMGB1诱导内皮细胞凋亡至关重要。     

槲皮素诱导人结肠癌Lovo细胞凋亡及其机制的研究

目的 目的 :探讨槲皮素诱导结肠癌Lovo细胞凋亡的机制.方法 结肠癌Lovo细胞在不同浓度槲皮素干预后分别采用流式细胞术、Western印迹法等检测细胞周期变化、凋亡及Bcl-2、Bax、p53及Caspase-3蛋白表达.结果 槲皮素作用于结肠癌Lovo细胞后,引起细胞凋亡增加,并出现G2/M期阻滞,Bcl-2蛋白表达抑制,p53、Bax及Caspase-3活性增强.结论 槲皮素可能通过激活p53、Bax及Caspase-3,抑制Bcl-2来诱导Lovo细胞凋亡.     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.     

Microbiology of human immunodeficiency virus anorectal disease

PURPOSE: Individuals who are seropositive for the human immunodeficiency virus are at high risk for opportunistic infection and anorectal disorders. Little prospective information is available regarding anorectal pathogens in these patients. METHODS: One hundred sixty-three HIV-seropositive patients presented to the colorectal clinic between 1989 and 1992. Forty-seven (29 percent) patients were thought to have an infectious process and were prospectively studied using a standardized multiculture protocol. RESULTS: Mean age was 33 (range, 19–59) years. All were male; high-risk behavior accounted for 87 percent of HIV transmissions. Presenting complaints included anorectal pain (79 percent), pus per anum (28 percent), and blood per anum (26 percent). Examination revealed perianal tenderness (60 percent), condyloma (38 percent), perianal ulcers (38 percent), and anal fissures (34 percent). Sixty-six sets of cultures were performed; 28 patients had one set, 15 had two sets, and 4 had three sets. Thirty-two of these 47 patients (68 percent) had positive cultures including herpes (50 percent), cytomegalovirus (25 percent),Neisseria gonorrhoeae (16 percent), chlamydia (16 percent), acidfast bacilli (2 percent), and others (9 percent). Six of 32 patients with positive cultures had more than one organism cultured. Sixteen (50 percent) patients with positive cultures were treated medically, 8 (25 percent) were treated surgically and 8 (25 percent) were treated with both modalities. Sixty-one procedures were performed on 17 patients for condylomata. Eighteen patients had 20 procedures for abscesses, 50 percent of whom had positive cultures for other than common bowel flora; all improved. Fourteen patients underwent 33 procedures for perianal fistulas.Mycobacterium fortuitum was cultured from one patient who required 13 procedures for abscesses and fistulas. Forty-five (96 percent) patients were followed for an average of 12.5 months ±2.9 SEM (range, 1–94 months). Symptoms were improved or resolved in 22 of 32 (69 percent) patients with positive cultures and in 11 of 13 (84 percent) with negative cultures. CONCLUSIONS: Specific pathogens may often be identified in human immunodeficiency virus-seropositive patients with anorectal disorders if aggressively sought. Although patients without specific pathogens identified may be expected to improve with planned empiric treatment, positive identification allows more directed therapy.