健脾消癌方通过PI3K/Akt信号通路抑制结肠癌血管生成的研究＊

目的 观察结肠癌HCT116细胞健脾消癌方的条件培养液对HUVEC细胞管腔形成的影响，从PI3K/Akt生物轴调控角度探讨其作用机制。方法 培养HCT116细胞，细胞设3组：对照组，健脾消癌方组（加入15%健脾消癌方含药血清）及人参皂苷Rg3组；制备HCT116细胞健脾消癌方条件培养液（分组及制备方法见实验方法），用条件培养液干预HUVEC（脐静脉内皮细胞，Human Umbilical Vein Endothelial Cells），Matrigel基质胶法检测HCT116细胞健脾消癌方条件培养液对HUVEC小管形成的影响。随后采用蛋白免疫印迹法（Western blot）检测各组HCT116细胞磷脂酰肌醇3-激酶（PI3K）、蛋白激酶B（Akt）、p-Akt、VEGF（血管内皮生长因子，Vascular endothelial growth factor）蛋白表达。最后在结肠癌HCT116荷瘤小鼠中验证健脾消癌方对肿瘤生长速度的影响，并经瘤组织VEGF蛋白表达、CD31免疫组化染色检测肿瘤内血管生成情况。结果 模型组HUVEC细胞管腔形成较空白血清组显著增加（P<0.05）；健脾消癌方组及人参皂苷Rg3组较模型组HUVEC细胞管腔形成显著减少（P<0.01）。p-Akt和VEGF蛋白表达水平模型组高于空白血清组（P<0.05），健脾消癌方组及人参皂苷Rg3组显著低于模型组（P<0.01）；PI3K、Akt蛋白表达量组间差异无统计学意义。与对照组比较，模型组荷瘤小鼠肿瘤体积显著性增大，瘤组织内VEGF表达、CD31阳性面积显著性增加，差异有统计学意义（P<0.05）；与模型组比较，健脾消癌方组及人参皂苷Rg3组荷瘤小鼠肿瘤体积显著减小，瘤组织内VEGF表达、CD31阳性面积降低，差异有统计学意义（P<0.05）。结论 健脾消癌方可抑制肿瘤的血管生成和生长，其作用机制可能与PI3K/Akt生物轴调控VEGF表达有关。     

健脾生血片治疗慢性心力衰竭伴贫血的临床效果

目的:探索健脾生血片治疗慢性心力衰竭伴贫血的疗效、安全性和作用机制。方法:选取2016年5月至2017年2月同济医院收治的慢性心力衰竭贫血患者144例,按照随机数字表法分为观察组和对照组,每组72例。观察组给予健脾生血片治疗,3片/次,3次/d,疗程3个月;对照组给予生血宝合剂治疗,15 m L/次,3次/d,3个月为1个疗程。比较2组患者治疗前与治疗后血红蛋白、红细胞计数、网织红细胞、血清铁、转铁蛋白饱和度、血清铁蛋白、血清铁调素(Hepcidin)、血清IL-1β、血清肿瘤坏死因子-α(TNF-α)、血清C反应蛋白(CRP)、左室射血分数(LVEF)、6 min步行距离、明尼苏达心力衰竭生命质量量表(MLHFQ)和不良事件。并随访2组心血管事件次数,住院次数与全因死亡率。结果:观察组72例患者完成了前3个月的治疗,随访期间脱失2例;对照组治疗期间1例患者退出研究,随访期间脱失4例。2组一般资料比较,差异无统计学意义(P 0. 05),具有可比性。观察组贫血有效率98. 6%,对照组有效率11. 3%,差异有统计学意义(P0. 05)。治疗后观察组红细胞计数和网织红细胞、均显著高于对照组(P 0. 05)。观察组血清铁、转铁蛋白饱和度水平均高于对照组,差异有统计学意义(P 0. 05),但血清铁调素水平显著低于对照组,差异有统计学意义(P 0. 05),血清铁蛋白水平2组差异无统计学意义(P 0. 05)。观察组IL-1β、血清TNF-α、血清CRP均显著低于对照组,差异有统计学意义(P 0. 05)。观察组LVEF、6 min步行距离、明尼苏达心力衰竭生命质量量表(MLHFQ)均显著高于对照组,差异有统计学意义(P 0. 05)。2组不良事件总发生率比较,差异无统计学意义(P 0. 05),但对照组4例患者出现血清肌酐、尿素氮水平异常,发生率高于观察组,差异有统计学意义(P 0. 05)。经1年随访,观察组心血管事件人均发生次数显著少于对照组(P 0. 05),但2组住院次数和全因死亡率比较,差异无统计学意义(P 0. 05)。结论:健脾生血片可有效治疗心力衰竭伴贫血,减少心血管发生次数,并且安全性良好,其作用机制与提供准确足量铁元素、抑制铁调素表达,抑制慢性炎性反应有关。     

益气复智颗粒对多发脑梗死性痴呆模型大鼠脑组织细胞凋亡的影响

目的 观察益气复智颗粒对多发脑梗死性痴呆模型大鼠脑皮质形态学、细胞凋亡的影响。方法 采用颈内动脉注射血栓的方法，复制多发梗死性痴呆大鼠模型，观察益气复智颗粒12．42g/kg分别于手术前、手术前后、手术后灌胃对实验动物脑皮质形态学、细胞凋亡的影响。结果 益气复智颗粒能使脑缺血后脑内神经细胞凋亡数目下降。结论 益气复智颗粒具有较好的保护脑神经元，阻断脑缺血致神经细胞死亡病理过程的作用。     

HPLC法测定滑膜炎颗粒中原儿茶醛的含量

目的建立滑膜炎颗粒剂中原儿茶醛的含量测定方法质量。方法采用高效液相色谱法对方中原儿茶醛进行含量测定。结果高效液相色谱法测定结果表明原儿茶醛在0．1005～1．005μg范围内呈线性关系，平均加样回收率为99．22％；RSD为0．72％。结论本法简便，重现性好、结果可靠，可作为控制滑膜炎颗粒剂的质量方法。     

Distribution of glutathione and glutathione-related enzyme systems in mitochondria and cytosol of cultured cerebellar astrocytes and granule cells

The cellular and regional distribution of glutathione (GSH) and GSH-related enzyme systems involved in cellular defense against reactive oxygen species and electrophilic xenobiotics in the nervous system has been extensively studied. However, little is known about the subcellular distribution of GSH systems in brain tissue and cultured neural cells. The present study investigates the distribution of mitochondrial and cytosolic GSH and GSH-related enzymes in cultured cerebellar astrocytes and granule cells, and compares them with levels in the adult rat cerebellum. Cytosolic GSH levels and cytosolic activities of glutathione reductase (GR), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) in astrocytes were 57, 153, 245, and 92% higher than those found in granule cells, respectively. In contrast, granule cells contained significantly higher mitochondrial GSH levels than astrocytes. Granule cells also demonstrated comparable mitochondria/cytosolic concentrations of GSH and GR, GPX and GST activities to those observed in the cerebellar tissue, whereas ratios in astrocytes were markedly lower. Although in vitro treatments with 100 μM ethacrynic acid depleted both cytosolic and mitochondrial GSH in cultured astrocytes and granule cells in a time-dependent fashion, cellular GSH in granule cells was more resistant to the GSH-depleting agent than astrocytes. These results suggest that although GSH and GSH-related enzymes are abundant in cytosolic compartments of astrocytes, mitochondrial pools are relatively small. Since brain mitochondria are sites of significant hydrogen peroxide generation, the mitochondrial localization of GSH and its associated enzymes in neural cells provide important defenses against toxic oxygen species in the nervous system. Differences in subcellular distribution of GSH systems in individual neural cell types may provide a basis for selective cellular and/or subcellular expression of neurotoxicity.     

扶正健脾汤配合环磷酰胺对荷瘤小鼠的实验研究

用环磷酰胺对荷瘤小鼠进行化疗同时配合扶正健脾汤进行治疗的疗效观察,证明扶正健脾汤具有减轻环磷酰胺的副作用,其表现为实验组小鼠的胸腺系数增大;脾内 IgG、IgM 阳性抗体形成细胞增多;肝内巨噬细胞的吞噬功能加强。提示扶正健脾汤具有拮抗环磷酰胺对骨髓、胸腺和脾脏的免疫抑制作用,为该方剂治疗肿瘤的临床应用提供实验依据。     

Ultrastructural pathology and cytochemical investigations of l-2-chloropropionic acid-induced neurointoxication of the rat cerebellum

The objectives of the studies described were to assess the ultrastructural neuropathology, blood-brain barrier (BBB) integrity and calcium status of the cerebellum of rats following a single dose of 750 mg · kg^–1 l-2-chloropropionic acid (l-2-CPA). The first indications of intoxication appeared at 36 h when condensation of many granule cells associated with Purkinje cell degeneration and marked astroglial swelling were observed. Some electron-lucent granule cells were also noted lying amongst these condensed forms. Condensed granule cells had swollen, electron-lucent mitochondria, dilated Golgi apparatus and nuclear crenation. Occasionally, areas of granule cell necrosis were also present at this time. Granule cell condensation probably represents a preliminary and irreversible stage in an excitotoxic process that leads to necrosis. At 48 and 72 h, most granule cells were necrotic, and occasionally, extravasation of both erythrocytes and leucocytes into the expanded extravascular space was observed. Evaluation of the BBB by ultrastructural cytochemical visualisation of horseradish peroxidase injected i.v. 2 min before killing by perfusion fixation showed substantial leakage. At 36 h post-dose, ultrastructural calcium localisation using oxalate/pyroantimonate precipitation demonstrated a substantial increase in calcium pyroantimonate precipitate in mitochondria and other membranous cytoplasmic organelles (especially the Golgi apparatus) in condensed granule cells, but with little in their nuclei. However, their immediate neighbours (of ostensibly normal ultrastructural appearances) contained greater amounts of intranuclear precipitate. Swollen astroglial cells (especially the Bergmann glia) contained considerable quantities of precipitate. A possible excitotoxic mechanism via l-2-CPA-induced NMDA receptor agonism leading to overwhelming calcium influx and disruption of cellular calcium homeostasis is proposed. Received: 8 May 1996 / Revised: 1 September 1996 / Accepted: 11 September 1996     

大鼠创伤性肠粘连模型的制备及黄厚排气颗粒对其粘连程度的影响

为筛选出操作方法简便，结果稳定可控的大鼠创伤性肠粘连模型，观察中药复方黄厚排气颗粒对其粘连程度的影响，选用SD大鼠，手术造成肠粘连模型，用中药复方2．5、5．0、10．0g/kg连续治疗7d后观察肠粘连程度。结果表明，以盲肠型动物模型的制备较为理想，手术易于操作，结果稳定可控。用中药复方治疗后模型动物的肠粘连程度得到明显改善。提示黄厚排气颗粒对术后肠粘连有一定的预防作用。     

养阴清热、宣痹通络法治疗类风湿性关节炎的临床研究

目的 观察清络通痹颗粒治疗类风湿性关节炎阴虚络热证的临床疗效。方法 将118例患者随机分为清络通痹颗粒治疗组63例和雷公藤多甙片对照组55例，观察用药前后主要症状、体征和实验室检测指标的变化。结果 治疗组的临床治愈率为9．52％，临床治愈显效率为38．10％，总有效率为90．48％，疗效明显优于对照组的0％、20％、83．64％（P＜0．05）；治疗组的类风湿因子滴度、血沉、C反应蛋白、免疫球蛋白也明显降低，与对照组相比差异显著（P＜0．05或P＜0．01）。结论 清络通痹颗粒具有抗炎镇痛、调节免疫紊乱等作用，从而达到控制病情的治疗效果。     

中药生精冲剂对精索静脉曲张大鼠的治疗

目的：研究中药生精冲剂对大鼠精索静脉曲张的影响及疗效。方法：从80只SD雄性大鼠中随机抽出20只作为假手术组，余60只均建立精索静脉曲张病理模型后随机均分为模型组、生精冲剂组和克罗米芬组。造模后15d生精冲剂组和克罗米酚组分别给予生精冲剂4g/（kg·d）和克罗米芬20mg/（kg·d）灌胃，模型组和假手术组正常喂食。造模后45d放免法测定血清性激素（FSH、LH和T）及观察各组大鼠睾丸组织结构。结果：生精冲剂组大鼠光镜下睾丸组织结构优于模型组和克罗米芬组；血清FSH、LH生精冲剂组显著低于模型组和克罗米芬组（P〈0.05），而克罗米芬组显著高于其他3组。T在生精冲剂组、克罗米芬组和假手术组之间无显著差异，但均显著高于模型组（P〈0.05）。结论：中药生精冲剂对精索静脉曲张引起的睾丸损害有保护及修复作用，且可能优于克罗米芬。