骨髓间充质干细胞移植治疗心力衰竭

目的 探讨骨髓间充质干细胞移植对阿霉素诱导的心力衰竭大鼠心功能的影响.方法 无菌条件下取8周龄F344大鼠的股骨和胫骨,获得骨髓间充质干细胞(MSCs),在体外纯化、扩增后 ,用5-溴-2'脱氧尿苷(BrdU)进行标记,然后注射到心力衰竭模型细胞移植组和对照组大鼠的心肌组织内,细胞移植后4周,采用生理记录仪测量3组大鼠的心功能 ,处死动物,心脏切片行免疫组化了解移植细胞在受体心脏的存活情况.结果 细胞移植后4周,细胞移植组大鼠死亡率为6.2%,明显低于假细胞移植组12.5%(P<0.01);在受体大鼠的心脏切片上有BrdU标记的移植细胞存活.心功能测定显示:与对照组相比,细胞移植组最大左室收缩末压(LVSP)、心率(HR)、左室内压最大(最小)变化速率(LV±dp/dtmax)均明显下降(P<0.01),而左室舒张末压(LVDP)明显升高(P<0.01);与对照组相比,细胞移植组大鼠的LVSP、HR、LV±dp/dtmax均有明显升高(P<0.01),而LVDP明显降低(P<0.01);与假移植组相比,细胞移植组大鼠的HR、 LV±dp/dtmax仍有明显下降(P<0.01),而LVDP明显升高(P<0.01).结论 MSCs移植可有效改善阿霉素诱导的扩张型心力衰竭大鼠的心功能,减少心力衰竭大鼠的死亡率.     

甲状腺素对心力衰竭大鼠心功能及心肌细胞钠-钙交换体表达的影响

目的:研究甲状腺素对心力衰竭大鼠的心功能及心肌细胞钠-钙交换体(NCX1)表达水平的影响。方法:SD 大鼠120只随机分为对照组(40只)、心力衰竭组(40只)和心力衰竭治疗组(40只)。通过缩窄主动脉复制心力衰竭动物模型,测定血流动力学指标,应用免疫组织化学及Western blot研究NCX1的表达水平。结果:(1)心力衰竭组左室收缩压(LVSP)和±dp/dtmax较对照组明显降低(P<0.01),甲状腺素治疗组LVSP和±dp/dtmax较对照组降低(P<0.05),但甲状腺素治疗组LVSP和±dp/dtmax较心力衰竭组明显升高(P<0.01);心力衰竭组左室舒张末压(LVEDP)较对照组明显升高(P<0.01),甲状腺素组LVEDP较心力衰竭组明显下降(P<0.01);(2)免疫组化单位面积(200倍视野/0.442mm2)内NCX1阳性颗粒数对照组(8.3±1.5)个,心力衰竭组为(24.1±4.5)个,甲状腺素治疗组为(15.6±2.9)个,各组间有显著性差异(P<0.01);(3)Western blot分析灰度峰值对照组13.7±2.4, 心力衰竭组为32.4±5.5,甲状腺素治疗组为20.2±3.9,各组间有显著性差异(P<0.001)。结论:甲状腺素可能通过抑制NCX1的过渡表达而发挥改善心力衰竭时心功能的作用。     

丹参素异丙酯对慢性缺氧大鼠血压和左心室功能的影响

目的: 研究丹参素异丙酯对缺氧大鼠左心室功能的影响。方法: 制备慢性缺氧大鼠模型,分别用大、小剂量的丹参素异丙酯灌胃,设立维拉帕米阳性对照组和正常对照组,采用大鼠心导管插入的方法,测定大鼠血流动力学和心功能指标,观察不同剂量丹参素异丙酯对慢性缺氧大鼠左室心功能的影响。结果: 与正常组比较,单纯模型组的动脉收缩压（SBP）、动脉舒张压（DBP）、平均动脉压（MBP）、左室收缩压（LVSP）、最大上升速率（+dp/dtmax）和最大下降速率（-dp/dtmax）显著升高（P<0.05或P<0.01）,T值(t-dp/dtmax)显著降低（P<0.05）,HE染色显示,心肌细胞出现肌纤维断裂、溶解,细胞核固缩、溶解。与单纯模型组比较,小剂量组的DBP、MBP、心率(HR)、左室舒张末压(LVEDP)和-dp/dtmax显著降低（P<0.05或P<0.01）,T值显著升高（P<0.05）;大剂量组的SBP、DBP、MBP、HR、LVEDP、左室舒张压(LVDP)、+dp/dtmax和-dp/dtmax显著降低（P<0.05或P<0.01）,T值显著升高（P<0.01）;HE染色显示,大、小剂量组心肌细胞内糖元含量丰富,肌纤维排列相对规则,未见明显的肌纤维断裂、溶解及核分裂。与阳性对照组比较,小剂量组的DBP、MBP、LVSP、LVDP和T值显著升高（P<0.05或P<0.01）,LVEDP显著降低（P<0.05）;大剂量组的DBP和T值均显著升高（P<0.05或P<0.01）,LVDP、LVEDP和+dp/dtmax均显著降低（P<0.05或P<0.01）。与小剂量组比较,大剂量组的LVDP、LVEDP和+dp/dtmax均显著降低（P<0.01）。结论: 丹参素异丙酯对慢性缺氧造成的大鼠血压升高、心率加快、心功能亢奋等有明显的抑制作用,因而对缺氧大鼠的心功能有明显的保护作用。     

Wistar大鼠慢性心力衰竭模型的制备及指标判定

目的使用阿霉素(ADR)制备Wistar大鼠慢性心衰模型,并对心功能指标进行检测,为研究心衰发病机制和探寻有效治疗方法提供理想的动物模型。方法将实验动物分为两组,一组是正常对照组,注射等体积生理盐水;另一组是实验组,采用阿霉素大鼠尾静脉注射法制备心衰模型,每周1次,共6 w。观察大鼠行为特征,末次注药2 w后称量大鼠终末体重(BW),心室重(VW),计算心体比(VW/BW),并进行心功能检测,包括心率(HR)、左室收缩压(LVSP)、左室舒张末压(LVEDP)及左室最大压力上升及下降速度(±dp/dtmax),并对心肌组织进行病理学检测。结果与正常对照组相比,实验组大鼠VW/BW明显增加,LVSP明显降低,LVEDP明显升高,±dp/dtmax则显著减小(P<0.05),病理学检测结果表明心肌出现典型慢性心衰样病理改变。结论连续多次尾静脉注射ADR可显著损伤心脏的舒缩功能,是建立大鼠心肌病及慢性充血性心力衰竭模型的一种简便可靠的方法。     

缬沙坦对心力衰竭心肌细胞收缩功能和钙瞬变的影响

目的:探讨缬沙坦对心力衰竭(HF)大鼠心肌细胞收缩功能和钙瞬变的作用。方法:结扎大鼠冠状动脉左前降支8周后,复制充血性HF模型,随机分为2组,即缬沙坦治疗组与HF对照组,分别用缬沙坦和安慰剂治疗,另设假手术组。治疗12周后,用急性酶解法获得单个大鼠心室肌细胞,用共聚焦显微镜测定单个细胞的收缩功能和钙瞬变。结果:①HF对照组左室舒张末压(LVEDP)明显大于假手术组(P<0.01),左室最大收缩压(LVSP)血压和左室内压的最大上升/下降速率(±dp/dtmax)明显小于假手术组(P<0.05、P<0.05和P<0.01);而缬沙坦治疗组LVSP和±dp/dtmax明显高于HF对照组(P<0.05和P<0.01),LVEDP明显小于HF对照组(P<0.01)。②HF对照组的心肌细胞表面积和最大舒张长度均大于假手术组(P<0.01),缩短分数明显低于假手术组(P<0.05),经缬沙坦治疗后均有明显改善(P<0.01)。③HF对照组心肌细胞的钙瞬变明显低于假手术组(P<0.01),舒张末期钙浓度明显升高(P<0.05),钙浓度下降时间明显减慢(P<0.01),经缬沙坦治疗后均明显改善(P<0.01)。结论:缬沙坦治疗能明显改善充血性HF的心肌细胞收缩功能,可能与改善HF心肌细胞钙调控异常有关。     

AMPK抑制剂P5499拮抗无钙预处理心肌保护作用

目的:观察腺苷酸活化的蛋白激酶(AMPK)抑制剂P5499对短暂无钙预处理心肌保护作用的影响。方法:对健康SD雄性大鼠心脏行Langendorff离体灌流,实验全程记录心脏冠脉流量(CF)、左心室内压(LVP)、左室舒张末压(LVEDP)、室内压最大变化速率(±dp/dtmax)及心率(HR),并计算左室发展压(LVDP)和心率-压力乘积(RPP)评价心功能的变化。再灌注结束后,采用2、3、5-氯化三苯基四氮唑(TTC)染色法评价心肌梗死(MI)面积的变化。结果:缺血/再灌注(I/R)后,心功能显著降低,CF明显减少(P<0.01),MI面积的比率为(39.6±1.49)%。短暂无钙预处理(CPC)可使LVEDP明显降低(P<0.05),CF、LVDP、±dp/dtmax及RPP均明显改善(P<0.01),MI面积显著缩小(P<0.01)。缺血前单独给予P5499对心功能、CF及MI面积无明显影响,但其可显著逆转CPC的心肌保护作用(P<0.01)。结论:AMPK可能是Ca2+预处理产生心肌保护信号的下游分子。     

骨髓间充质干细胞移植对心力衰竭大鼠心功能的影响

目的探讨骨髓间充质干细胞（MSCs）对心衰大鼠心功能的影响及其机制。方法腹腔注射阿霉素（2mg／kg,每周1次,连续6周）至近交系F344大鼠体内,建立心衰大鼠模型。存活大鼠（32只）随机分为2组：心衰组（16只）和细胞移植组（16只）。分离纯化大鼠MSCs进行体外培养,予4,6-联脒-2-苯基吲哚（DAPI）进行标记;经股静脉注射MSCs或DMEM至心衰大鼠体内。细胞移植后4周,应用多导生理记录仪测量大鼠心功能;通过免疫组织荧光及化学染色,观察移植细胞存活情况,并计算血管数量;通过天狼猩红染色计算心肌胶原容积分数（CVF）和血管周围胶原面积（PVCA）。结果移植后4周,细胞移植组大鼠心肌组织见到DAPI标记的移植细胞存活,并表达心肌特异性抗原心肌肌球蛋白重链（β—MHC）。与心衰组相比,细胞移植组最大左室收缩末压（LVSP）、左室内压最大（最小）变化速率（LV＋dp／dtmax）均明显升高（P〈0．05）,而左室舒张末压（LVDP）明显下降（P〈0．05）;细胞移植组左室血管数量明显高于心衰组[（11．83±1．40）个比（7．78±1．39）个,P〈O．05],细胞移植组左心室内膜CVF及PVCA明显低于心衰组（4．53±1．98比7．79±1．99,10．91±2．31比14．17±2．49,P〈0．05）。结论MSCs移植可改善心衰大鼠心功能,其机制可能与MSCs归巢至心脏,分化为心肌样细胞,并且促进血管新生、抑制心肌纤维化有关。     

心脏离体灌流装置观察细胞移植对心脏功能的影响

目的观察胚胎心肌细胞移植到大鼠急性心肌梗死区后对大鼠心脏功能的影响及其存活情况。方法100只成年大鼠随机分成2组。移植组开胸结扎大鼠冠状动脉后在梗死区内注射培养的胚胎心肌细胞悬液;对照组结扎冠状动脉后在与移植组相同的部位注射等量的细胞培养液。各组分别在移植后的48h、72h、1周、2周收集心脏标本,HE染色及抗5鄄溴脱氧尿核苷(5鄄BrdU)免疫组化染色。Langendorff心脏离体灌流装置测定移植术后第2周各组大鼠的心脏功能。结果移植组大鼠心脏左室收缩峰压(LVSP)、左室内压最大上升与下降速率( dp/dtmax,-dp/dtmax)与对照组相比明显升高,差异有非常显著意义(P<0.01),左室舒张期末压(LVEDP)明显降低,与对照组相比差异也有非常显著意义(P<0.01)。在移植组心肌梗死区内可以看到存活的胚胎心肌细胞,对照组未发现。结论胚胎心肌细胞移植能够改善大鼠的心脏功能,并且可以在大鼠的心肌梗死区内存活及分化。     

糖尿病树鼩血栓性脑缺血时心脏血流动力学及心肌超微结构的变化

目的观察实验性糖尿病树鼩在血栓性局部脑缺血时心脏血流动力学及心肌超微结构的变化。方法链脲佐菌素(STZ)55mg/kg空腹腹腔注射制备糖尿病树鼩模型,一周后通过光化学反应诱导局部血栓性脑缺血。将树鼩随机分为糖尿病组,脑缺血组,糖尿病加脑缺血组,并设假手术对照组,每组8只,用多导生理仪观察左心室内压(LVSP)、左心室舒张末压(LVEDP),左心室收缩与舒张瞬间变化最大速率(±dp/dtmax),收缩动脉压(SAP),舒张动脉压(DAP)及心率(HR)。并取左心室心肌组织,行透射电镜观察心肌超微结构变化。结果 与假手术对照组相比,脑缺血组LVSP、+dp/dtmax下降(P<0.05),-dp/dtmax明显下降(P<0.01),LVEDP升高(P<0.05);糖尿病组-dp/dtmax下降(P<0.05),LVEDP明显升高(P<0.01)。与脑缺血组相比,糖尿病加脑缺血组LVSP、+dp/dtmax降(P<0.05),-dp/dtmax明显下降(P<0.01),LVEDP升高(P<0.05)。脑缺血组和糖尿病组心肌超微结构观察显示:心肌肌原纤维间水肿,一些肌原纤维可见收缩小带形成,部份线粒体固缩,部份线粒体肿胀。糖尿病加脑缺血组树鼩可见:大部分线粒体固缩,部分线粒体肿胀、嵴溶解,心肌纤维较多收缩带形成,间质充血水肿,毛细血管内皮细胞见较多空泡。结论树鼩血栓性脑缺血时伴有心脏血流动力学及心肌超微结构的异常,糖尿病急     

温补心阳和肾阳方对心力衰竭大鼠心功能和心肌磷酸腺苷含量的影响

目的研究温补心阳和肾阳方对心力衰竭大鼠心功能和心肌磷酸腺苷含量的影响。方法采用结扎冠状动脉前降支术制备心力衰竭大鼠模型,用温补心阳和温补肾阳方于术后2周灌胃,以假手术组为空白对照组,曲美他嗪为阳性药物对照组。给药8周,采用酶联免疫吸附法检测心梗后心力衰竭大鼠药物干预前后脑钠肽(BNP)水平,测定治疗后血流动力学改变,包括心率(HR)、左室收缩压(LVSP)、左室舒张压(LVEDP)、左室内压上升下降最大速率(±LVdp/dtmax),并行高效液相法测定心肌组织中AMP、ADP、ATP的含量。结果对心力衰竭大鼠心功能的影响,温补心阳和肾阳组较模型组,显著降低BNP水平(P<0.01),改善心肌收缩、舒张功能。而肾阳组大鼠LVSP、+LVdp/dtmax较心阳组升高(P<0.05),LVEDP、-LVdp/dtmax、HR、BNP较心阳组降低(P<0.05)。对心力衰竭大鼠心肌磷酸腺苷含量的影响:温补心阳和肾阳组大鼠较模型组心肌AMP、ADP降低(P<0.05),ATP含量升高,而肾阳组大鼠心肌ATP含量显著高于模型组(P<0.01),较心阳组更接近曲美他嗪和假手术组。结论温补心阳和温补肾阳可改善心力衰竭大鼠能量代谢,提高心功能,其中以温补肾阳作用更显著。     

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.     

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.     

N-acetyl-L-cysteine combined with mesalamine in the treatment of ulcerative colitis： Randomized,placebo-controlled pilot study

AIM： To evaluate the effectiveness and safety of oral N-acetyl-L-cysteine （NAC） co-administration with mesalamine in ulcerative colitis （UC） patients.
METHODS： Thirty seven patients with mild to moderate UC were randomized to receive a four-wk course of oral mesalamine （2.4 g/d） plus N-acetyl-L-cysteine （0.8 g/d） （group A） or mesalamine plus placebo （group B）. Patients were monitored using the Modified Truelove-Witts Severity Index （MTWSI）. The primary endpoint was clinical remission （MTWSI ≤ 2） at 4 wk. Secondary endpoints were clinical response （defined as a reduction from baseline in the MTWSI of ≥ 2 points） and drug safety. The serum TNF-α, interleukin-6, interleukin-8 and MCP-1 were evaluated at baseline and at 4 wk of treatment. RESULTS： Analysis per-protocol criteria showed clinical remission rates of 63% and 50% after 4 wk treatment with mesalamine plus N-acetyl-L-cysteine （group A） and mesalamine plus placebo （group B） respectively （OR = 1.71; 95% CI： 0.46 to 6.36; P = 0.19; NNT = 7.7）. Analysis of variance （ANOVA） of data indicated a significant reduction of MTWSI in group A （P = 0.046） with respect to basal condition without significant changes in the group B （P = 0.735） during treatment. Clinical responses were 66% （group A） vs 44% （group B） after 4 wk of treatment （OR = 2.5; 95% CI： 0.64 to 9.65; P = 0.11; NNT = 4.5）. Clinical improvement in group A correlated with a decrease of IL-8 and MCP-1. Rates of adverse events did not differ significantly between both groups.
CONCLUSION： In group A （oral NAC combined with mesalamine） contrarily to group B （mesalamine alone）, the clinical improvement correlates with a decrease of chemokines such as MCP-1 and IL-8. NAC addition not produced any side effects.     

Delorme's transrectal excision for internal rectal prolapse

Surgical therapy of functional outlet obstruction in patients with internal rectal intussusception may include abdominal, perineal, or transrectal procedures. Because abdominal procedures often result in significant physiologic impact but unrelieved constipation, the authors have elected Delorme's transrectal excision for management of these patients. Since a short-term placebo effect attends many therapies, this report describes results of transrectal excision only after a threeyear postoperative period. Delorme's transrectal excision of internal intussusception accomplished sustained symptomatic relief in over 70 percent of otherwise refractory constipated patients. The association of internal intussusception with other abnormalities underscores the importance of defining both anatomic and functional components when selecting patients whose constipation may require surgical therapy. Critical technical elements, surgical pitfalls, and potential complications of the procedure are discussed.Poster presentation at the meeting of The American Society of Colon and Rectal Surgeons, Toronto, Canada, June 11 to 16, 1989.     

The oral glucose tolerance test: A comparison of the time points on the basis of limit values,normal dispersion,and reproducibility

Summary Time points in the glucose tolerance test (GTT) are compared on the basis of limit values, dispersion within a reference population, and reproducibility. We suggest using the distance between a limit value and the median reference value as a measure of the magnitude of abnormality. The distance between 140 mg/100 ml and the median fasting plasma glucose value is chosen as a standard distance and limits for other points in the GTT are calculated to equal this standard distance of abnormality. We suggest that the probability of correctly interpreting an inividual result is directly related to the reproducibility of the test and inversely related to the percentage of the total range of values which is dispersed among the normal population. The ratio of reproducibility to percentage normal dispersion is proposed as an index of the probability of correctly interpreting an individual result. According to this index, the probability of correct interpretation varies in order: fasting plasma glucose concentration>3-h>2-h>0.5-h>1-h plasma glucose concentration.