移植时机对骨髓单个核细胞治疗心肌梗死效果的初步研究

目的：研究经冠状动脉移植骨髓单个核细胞治疗心肌梗死的最佳时间窗。方法：结扎小型猪冠状动脉前降支90 min 再灌注制备心肌梗死模型,分为2小时移植组、2小时对照组、1周移植组和1周对照组,每组7只。移植组在结扎后2小时和1周经冠状动脉移植(8～10)×10~7个 PKH26标记的骨髓单个核细胞,对照组注射等体积培养基结扎后6周检测心功能变化并行了病理检查。结果：两个移植组在心肌梗死区均可找到移植细胞,其Ⅷ因子和 desmin 免疫组化染色均为阳性。血流动力学指标显示：结扎后6周较结扎后90分钟相比.2小时移植组的左心室压力最大下降速度和心排量显著升高(P＜0.05)；1周移植组的左心室舒张未压和心排量显著改善(P＜0.05)。超声心动图指标显示结扎后6周时2小时移植组的每搏排血量和心排量显著高于2小时对照组(P＜0.05)；1周移植绀结扎后6周-结扎前射血分数的差值明显高于1周对照组 (P＜0.05)。两移植组结扎后6周的各项指标均无显著差异(P 均＞0.05)。1周移植组梗死区的小血管数量明显高于2 小时移植组(P＜0.01)。结论：在结扎后2小时和1周时经冠状动脉移植的骨髓细胞均能定植于心肌梗死区,并可向心肌细胞和血管内皮细胞方向分化,有改善心功能的潜能。在结扎后1周移植能更有效地促进小血管新生。     

经冠状动脉注射骨髓单个核细胞影响心肌细胞凋亡的实验研究

目的观察经冠状动脉注射方式移植骨髓单个核细胞对心脏功能的改善和心肌细胞凋亡的影响。方法结扎小型猪冠状动脉制备心肌梗死模型，然后经冠状动脉注射骨髓单个核细胞，术后3周用超声心动图以及左心室造影检测心功能．核素心肌显像观察心肌灌注，冠状动脉造影观察侧支循环形成．用TUNEL检测心肌细胞凋亡。结果把骨髓单个核细胞通过冠状动脉注射到猪心肌梗死模型，显示左心室dp／dtmax较对照组增高，心肌灌注显著改善。冠状动脉有侧支循环形成，缺血心肌细胞；较对照组减少了53．6％结论骨髓单个核细胞心肌内移植可改善心脏收缩功能，可能与移植细胞抑制心肌细胞凋亡有关。     

经冠状动脉注射骨髓单个核细胞治疗缺血性心肌损伤的实验研究

目的　证实骨髓干细胞心肌内移植治疗对心脏功能的改善和促血管新生作用。方法　小型猪冠状动脉结扎制备心肌梗死模型 ,然后经冠状动脉内注射骨髓单个核细胞 ,术后 3周用超声心动图以及左心室造影检测心功能的变化 ,核素心肌显像观察心肌灌注的情况 ,冠状动脉造影观察侧支循环的形成 ,免疫组化计数血管密度。结果　心肌梗死小型猪冠状动脉注射骨髓单个核细胞后 ,左心室造影显示左室 dP dtmax与对照组比较增高。核素心肌显像显示冠状动脉注射骨髓单个核细胞后心肌灌注显著改善。移植治疗后冠状动脉造影显示有侧支循环形成 ,血管密度计数比对照组增加了5 2 2 % (5 6 6± 11 7 mm2 vs 37 2± 8 4 mm2 ,P <0 0 5 )。结论　骨髓单个核细胞心肌内移植可能通过促进血管新生改善心脏收缩功能。     

骨髓动员与细胞移植对直接经皮腔内冠状动脉成形术后急性心肌梗死治疗作用

目的比较骨髓间充质干细胞移植和骨髓动员对急性心肌梗死再通模型的心脏修复作用.方法人重组粒细胞集落刺激因子静注1d,皮下注射连续5d行骨髓动员,骨髓间充质干细胞(4～8)×106经梗死相关冠状动脉注入行细胞移植.3周后观察超声心动图指标;免疫组织化学染色行不同区域血管计数,并观察增殖期内皮细胞和心肌细胞或再生心肌细胞.结果骨髓动员组和细胞移植组心功能明显改善;二组均可见心肌细胞再生;细胞移植组和骨髓动员组均可见大量新生血管,但二者分布区域不同.结论骨髓动员及骨髓间充质干细胞移植均可明显改善心肌梗死后心功能,二者可促进心肌细胞再生且对不同区域的血管再生作用不同.     

骨髓干细胞移植可影响心肌梗死后心肌细胞的凋亡

目的观察骨髓单个核细胞（bone marrowmononuclear cells,BM-MNCs）移植对大鼠急性心肌梗死（AMI）后心肌细胞凋亡的影响。方法健康雄性Wistar大鼠30只,随机均分为实验组和对照组。用结扎冠状动脉左前降支的方法建立大鼠AMI模型,将制备的BM-MNCs悬液和培养液分别经心外膜下植入实验组和对照组梗死心肌周围。移植术后4周末,检测心肌细胞凋亡情况及Bcl-2、Fas、FasL蛋白在心肌细胞中表达水平,并观察梗死区心肌内移植BM-MNCs及其周边区组织形态学特点。结果移植后4周末,实验组心肌细胞凋亡指数显著低于对照组（P<0.05）;与对照组相比,实验组Bcl-2蛋白吸光值显著升高（P<0.05）,Fas、FasL蛋白吸光值显著降低（P<0.05）;实验组心肌梗死区内有BrdU标记阳性的BM-MNCs存活。结论同种异体大鼠BM-MNCs移植可调节Bcl-2、Fas、FasL蛋白表达,抑制AMI后心肌细胞凋亡发生。     

自体骨髓间充质干细胞移植对急性心肌梗死后心功能的影响

目的观察骨髓间充质干细胞（MSCs）经冠脉移植对急性心肌梗死后心功能的影响。方法24只日本大耳白兔,随机分为MSCs移植组（n=12）和培养液对照组（n=12）。从兔股骨抽取骨髓,体外培养MSCs。通过结扎左冠前降支建立急性心肌梗死模型。冠脉结扎后7d,细胞移植组和对照组直接经冠脉注入MSCs和培养液。于心肌梗死前、细胞移植前、细胞移植后1、2和4周对兔进行超声心动图检查。移植后4周处死动物,进行BrdU和第Ⅷ因子相关抗原免疫组化检测。结果移植后2周,MSCs移植组在射血分数（LVEF）和左室收缩末直径（LVESD）方面与移植前和对照组相比有显著改善（P<0.05）;移植4周后,LVEF、LVESD和左室舒张末直径（LVEDD）在MSCs移植组与移植前及对照组相比均有显著改善（P<0.05）。免疫组化检测发现,MSCs移植组BrdU染色阳性,血管计数较对照组明显增多（P<0.01）。结论经冠脉移植的MSCs可在梗死区心肌内存活并逐渐分化成心肌样细胞,促进毛细血管生成,显著改善心功能。     

兔骨髓间充质干细胞自体移植及血管新生修复梗死心肌的实验研究

目的　了解骨髓间充质干细胞 (MSCs)在心肌梗死的缺血心肌微环境中转化为心肌细胞并改善心功能的可行性。方法　结扎兔左前降支 (LAD)制作急性心肌梗死(AMI)模型 ,骨穿抽取骨髓 ,分离培养扩增骨髓间充质干细胞。术后 2周进行心肌梗死瘢痕内移植 5溴脱氧尿嘧啶(BrdU)标记的骨髓间充质干细胞 ,对照组注射培养基。AMI术前、AMI术后 3d及移植后 4周做超声心动图检测心功能 ,移植后 4周从右颈总动脉插管至左心室测左室压力变化 ,随后处死动物模型 ,取左室标本做冰冻切片 ,采用免疫荧光方法检测BrdU鉴定植入细胞并检测Ⅷ因子相关抗原以测量毛细血管密度。结果　细胞移植组左室切片中可见BrdU染色阳性细胞即植入细胞 ,对照组未见BrdU染色阳性细胞。Ⅷ因子阳性内皮细胞计数表明细胞移植组毛细血管密度大于对照组 (P <0 0 5 )。超声心动图检查证实MSCs移植后 4周左室收缩末期直径 (LVESD)、舒张末期直径 (LVEDD)均小于对照组 (分别为P <0 0 1及P <0 0 5 ) ,小轴缩短率 (ΔD % )、射血分数 (EF)均大于对照组 (均P <0 0 1)。结论　兔缺血心肌内注射骨髓间充质干细胞可依赖组织微环境转化为心肌细胞修复梗死心肌 ,可能伴有移植区域新生血管形成 ,能显著改善心功能 ,可用于心肌梗死的治疗。     

梗死心肌血管微环境对经冠状动脉内移植骨髓干细胞疗效的影响

目的观察梗死区侧支循环对冠状动脉(冠脉)内骨髓干细胞移植术心肌修复疗效的影响。方法结扎猪左前降支建立心肌梗死模型,2周后行冠状动脉造影进行梗死区侧支循环Rentrop评分,取分值为0(R0)或1(R1)的动物各10头,经冠脉内注射骨髓单个核细胞(BMMNCs)或等体积磷酸盐缓冲液(PBS,对照组),随机分为R0 BMT,R0 PBS,R1 单个核细胞移植(BMT)和R1 PBS4组(n=5)。结果梗死后6周,R1 BMT组Rentrop积分显著升高;R0 BMT与R1 BMT两组心功能均明显提高,以R1 BMT组最显著;血管新生和移植细胞数量在R1 BMT组最明显。结论冠脉内移植BMMNCs对梗死心肌有修复作用,而梗死区侧支循环的存在放大其疗效。     

经冠脉移植骨髓单个核细胞和间充质干细胞治疗急性心肌梗死的对比实验研究

目的 对比研究经冠脉骨髓单个核细胞（BM-MNC）和间充质干细胞（MSC）移植对实验性急性心肌梗死（AMI）心功能的影响及其机制.方法 选用12只雄性冀中白猪随机分为：正常对照组、AMI模型组、BM-MNC组、MSC组各3只,经导管球囊封闭前降支制作AMI的动物模型,于梗死后1 h直接冠脉球囊成型术后经OTW球囊注入骨髓干细胞.分别于术前及术后4 w经心脏超声检测心功能,4 w后取材行光、电镜病理学检查,实时定量RT-PCR检测心肌血管内皮生长因子（VEGF）和碱性成纤维细胞生长因子（bFGF）mRNA表达.结果 4 w时干细胞组比AMI模型组室壁运动异常指数显著减轻（P＜0.05）、射血分数显著提高（P＜0.01）.与AMI模型组相比：BM-M;NC和MSC组梗死区及梗死边缘区血管数显著增多、BM-MNC组增加比MSC组显著（P＜0.01）,心肌细胞凋亡指数显著降低,BM-MNC组及MSC组间无明显差异（P＜0.01）.干细胞移植组梗死边缘区冠脉血管周围可见异常细胞生长,有毛细血管＂芽生＂现象,可见不成熟的心肌细胞和细胞凋亡.4 w时左室射血分数（LVEF）与心肌血管数成正相关（r=0.694 9,P=0.037 7）,LVEF与心肌细胞凋亡指数成负相关（r=0.913 3,P=0.000 6）.BM-MNC组,心肌梗死区及梗死边缘区VEGF基因表达比其他三组均明显增加（梗死区F=4.23,P=0.045 6,边缘区F=5.66,P=0.022 3）.BM-MNC及MSC组心肌梗死区bFGF基因表达比梗死模型组显著增加（梗死区F=7.49,P=0.010 4）.结论 经冠脉骨髓单个核细胞和间充质干细胞移植均可改善实验性AMI心功能;改善心功能的机理与梗死区及梗死边缘区VEGF及bFGF表达增加,血管密度增加,心肌细胞凋亡减少有关;骨髓单个核细胞移植的促血管增生作用优于间充质干细胞移植.     

冠状动脉内自体骨髓单个核细胞移植治疗急性心肌梗死患者的疗效

目的:探讨急性心肌梗死患者自体骨髓单个核细胞经冠状动脉移植的安全性和对心功能的保护作用。方法:2003年3月以来,84例急性心肌梗死患者急诊静脉溶栓或急诊经皮冠状动脉成形术(PTCA)加支架治疗后2周内行择期冠状动脉造影或PTCA加支架治疗,其中50例抽取骨髓40ml,提取单个核细胞,经冠状动脉注入,另34例不做自体骨髓单个核细胞经冠状动脉移植,作为对照组。81例患者术前和术后6个月、2年均行多巴酚丁胺负荷试验(另3例未完成超声心动图随访观察)。结果:治疗组患者临床随访无明显不良反应,心功能明显改善,运动耐量增加。多巴酚丁胺负荷试验左室射血分数(LVEF)和室壁运动记分指数(WMSI)显著改善,峰值LVEF和WMSI与基础状态LVEF和WMSI的差值在治疗前后相比,与对照组相比均差异有统计学意义。结论:自体骨髓单个核细胞经冠状动脉移植治疗急性心肌梗死患者,经6个月～2年的临床观察无明显不良反应,具备安全性,多巴酚丁胺负荷试验显示出自体骨髓单个核细胞治疗对梗死后心功能有保护作用。     

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.