应用国产微机导纳血流图仪监测健康人脑血流量的初步报告

利用导纳血流图仪对 91例健康人行双侧脑血流检测,研究不同年龄、不同性别健康人双侧脑血流导纳指标改变及其意义.     

Anticancer effect of realgar nanoparticles on mouse melanoma skin cancer in vivo via transdermal drug delivery

Realgar has been used successfully to treat diseases for thousands of years, but its poor water solubility and high toxicity hampered its further medical uses. Here, we first applied transdermal drug delivery system to deliver realgar nanoparticles to investigate its anticancer effect and toxicity in vivo. In this study, MTT assay and flow cytometry analysis demonstrated that realgar significantly suppressed the proliferation and induced apoptosis of B16 melanoma cells in a dose-dependent manner. Transdermal penetration studies in vitro showed realgar nanoparticles could be delivered efficiently through skin. Tests on tumor-bearing C57BL/6 mice displayed that realgar could decrease the tumor volume markedly via transdermal drug delivery compared with the intraperitoneal administration and the control. Hematoxylin–eosin and immunohistochemical staining revealed that it could inhibit angiogenesis. The monitoring of the hepatic injury, body weight, feeding behavior, motor activity, and skin irritation of each animal indicated little toxicity of realgar to mice. The results demonstrated that realgar nanoparticles can be dermally delivered to achieve high efficacy against menaloma in vivo with low toxicity.     

Blood and urinary metabolomic evidence validating traditional Chinese medicine diagnostic classification of major depressive disorder

Background

Major depressive disorder (MDD) is a highly heterogeneous disease. Further classification may characterize its heterogeneity. The purpose of this study was to examine whether metabolomic variables could differentiate traditional Chinese medicine (TCM) diagnostic subtypes of MDD.

Methods

Fifty medication-free patients who were experiencing a recurrent depressive episode were classified into Liver Qi Stagnation (LQS, n?=?30) and Heart and Spleen Deficiency (HSD, n?=?20) subtypes according to TCM diagnosis. Healthy volunteers (n?=?28) were included as controls. Gas chromatography-mass spectrometry (GC–MS) was used to examine serum and urinary metabolomic profiles.

Results

Twenty-eight metabolites were identified for good separations between TCM subtypes and healthy controls in serum samples. Both TCM subtypes had similar profiles in proteinogenic branched-chain amino acids (BCAAs) (valine, leucine, and isoleucine) and energy metabolism-related metabolites that were differentiated from healthy controls. The LQS subtype additionally differed from healthy controls in multiple amino acid metabolites that are involved in biosynthesis of monoamine and amino acid neurotransmitters, including phenylalanine, 3-hydroxybutric acid, o-tyrosine, glycine, l-tryptophan, and N-acetyl-l-aspartic acid. Threonic acid, methionine, stearic acid, and isobutyric acid are differentially associated with the two subtypes.

Conclusions

While both TCM subtypes are associated with aberrant BCAA and energy metabolism, the LQS subtype may represent an MDD subpopulation characterized by abnormalities in the biosynthesis of monoamine and amino acid neurotransmitters and closer associations with stress-related pathophysiology. The metabolites differentially associated with the two subtypes are promising biomarkers for predicting TCM subtype-specific antidepressant response [registered at http://www.clinicaltrials.gov (NCT02346682) on January 27, 2015].

     

血管生成素在COS-7细胞中的表达及其生物活性研究

本研究旨在实现血管生成素（ANG）在真核细胞中的表达并探讨其生物学功能。通过RT—PCR获得ang基因，构建真核表达载体pcDNA3．1-ang，在转染COS-7细胞后进行瞬时表达，通过Western blot对表达产物进行鉴定。利用MTT法分析表达上清对ECV304细胞的促增殖作用，同时利用鸡胚分析表达上清的促血管生成作用。结果表明：瞬时转染后细胞培养上清中有重组ANG的表达，并能与抗-ANG单克隆抗体发生特异性反应。与转染空载体的对照相比．转染pcDNA3．1-ang的细胞培养上清具有促进ECV304细胞增殖的作用，并能显著促进鸡胚尿囊膜血管生长。结论：ang可在COS-7细胞中瞬时表达，其表达产物具有显著的促细胞增殖和促进血管生成的作用。     

小鼠调节性树突状细胞分泌的外泌体诱导免疫耐受的实验研究

本研究探讨小鼠调节性树突状细胞（rDC）分泌的外泌体（regulatory exosomes，rDex）诱导免疫耐受的作用，并与正常未成熟树突状细胞的外泌体（immature exosomes，iDex）诱导免疫耐受的作用进行比较。取C57BL／6（H-2^b）小鼠骨髓细胞诱导未成熟树突状细胞（iDC），TGF—β1联合IL-10诱导调节性树突状细胞（rDC），流式细胞术检测两种DC的表型。采用超速离心结合膜超滤的方法分别提取rDex和iDex。建立小鼠皮肤移植模型．以C57BL／6小鼠为供者，以BALB／c（H-2^d）小鼠为受者，按对受者处理的不同实验分3组，iDex组术前7、3天受者经尾静脉注射10μg供者的iDex，rDex组术前7、3天受者经尾静脉注射10μg供者的rDex，另设PBS对照组，观察移植皮肤存活情况。通过单向混合淋巴细胞反应（MLR）观察供者及非亲缘异基因供者DBA／2对同种异基因小鼠T细胞增殖情况。结果表明：TGF—β1、IL-10可下调DC表面共刺激分子CD80、CD86、CD40的表达。移植皮片平均存活时间（MST），在对照组为7、8天，iDex组为10．7天，rDex组为18．8天；iDex组的移植皮片MST明显长于对照组（P〈0．05）；rDex组的移植皮片MST明显长于iDex组（P〈0．01）。MLR结果证明iDex组和rDex组B／C小鼠均对C57小鼠的脾细胞产生特异性耐受，尤其是rDex组；对非亲缘异基因DBA供者的脾细胞两组仍表现出强烈的免疫应答。结论：iDex和rDex均有诱导免疫耐受的作用，且rDex作用较iDex作用好。     

Formaldehyde induces neurotoxicity to PC12 cells involving inhibition of paraoxonase-1 expression and activity

1. Formaldehyde (FA) has been found to cause toxicity to neurons. However, its neurotoxic mechanisms have not yet been clarified. Increasing evidence has shown that oxidative damage is one of the most critical effects of formaldehyde exposure. Paraoxonase-1 (PON-1) is a pivotal endogenous anti-oxidant. Thus, we hypothesized that FA-mediated downregulation of PON1 is associated with its neurotoxicity. 2. In the present work, we used PC12 cells to study the neurotoxicity of FA and explore whether PON-1 is implicated in FA-induced neurotoxicity. 3. We found that FA has potent cytotoxic and apoptotic effects on PC12 cells. FA induces an accumulation of intracellular reactive oxygen species along with downregulation of Bcl-2 expression, as well as increased cytochrome c release. FA significantly suppressed the expression and activity of PON-1 in PC12 cells. Furthermore, H(2)S, an endogenous anti-oxidant gas, antagonizes FA-induced cytotoxicity as well as 2-hydroxyquinoline, a specific inhibitor of PON-1, which also induces cytotoxicity to PC12 cells. 4. The results of the present study provide, for the first time, evidence that the inhibitory effect on PON-1 expression and activity is involved in the neurotoxicity of FA, and suggest a promising role of PON-1 as a novel therapeutic strategy for FA-mediated toxicity.     

细菌纤维素构建组织工程角膜基质的方法及其评价

目的：探讨以细菌纤维素(BC)为支架构建组织工程角膜基质的可行性。方法： 体外分离培养兔和人角膜基质细胞,种植到BC膜中,构建完成后进行兔和人角膜基质细胞-BC复合膜的生物检测,并将兔角膜细胞生物复合物进行同种异体移植。术后1、4和8周时分别活体行前节OCT、角膜共焦显微镜检查,离体后组织学及免疫组织化学检查。结果： 人和兔角膜基质细胞长入BC的网架结构,细胞生长状态良好。移植术后1周兔眼无炎症反应及新生血管,4周后植片边缘出现新生血管,表面无水肿及溃疡形成。术后8周角膜共焦显微镜显示：移植片周围基质细胞增生活跃,邻近的角膜内皮细胞数量和形态正常。前节OCT显示：移植后复合膜逐渐降解,4周时复合膜边界模糊,8周时密度接近正常角膜组织。离体组织学检查显示：BC复合膜与正常角膜基质相似,有炎细胞浸润和血管形成,角膜组织无坏死和溶解。结论： BC无细胞毒性,具有良好的组织相容性和可降解性,可作为构建组织工程角膜基质的生物支架。     

基于生物检测的双黄连片溶出度研究(英文)

中药固体制剂的溶出度研究一直是中药现代化的难点问题。中药与化学合成药不同,中药药效物质十分复杂,临床疗效可能取决于其中的未知成分,也可能取决于各个成分的协同作用。单独使用化学方法测定中药固体制剂的溶出度不能全面反映其内在质量,本课题组首次提出基于生物检测的一种新的溶出度评价方法。在试验中,以清热解毒类中药固体制剂双黄连片为模型药,进行了溶出度研究。在pH值为6.8的溶解介质,即磷酸盐缓冲液中双黄连片不同溶出时间的溶液对金黄色葡萄球菌的抑制作用不同,应用微量量热法获得金黄色葡萄球菌的生长功率–时间曲线和该抑制作用的一系列生物热动力学参数。结果表明,基于生物热动力学的方法可以用于控制中药固体制剂的内在质量。生物热活性检测有望成为中药固体制剂体外溶出度评价的重要手段之一。     

Effects of a Shuangling Fuzheng anticancer preparation on the proliferation of SGC-7901 cells and immune function in a cyclophosphamide-treated murine model

AIM: To study the inhibitory effects of a Shuangling Fuzheng anticancer preparation (SFAP) on the human gastric cancer cell line SGC-7901 in vitro as well as its immune-modulated effects in a cyclophosphamide-treated murine model. METHODS: MTT experiments and immunocytochemistry ABC experiments were performed for detecting the proliferation of SGC-7901 cells in vitro and protein expression of c-myc. The staphylococcal protein A (SPA) rosette test was utilized for measuring the ratio of T-lymphocyte subsets from peripheral blood in a cyclophosphamide-treated murine model. Enzyme- linked immunosorbant assay (ELISA) was performed for measuring the levels of serum sIL-2R in treated mice, while immunoturbidimetry was used for measuring the levels of immunoglobulins (Ig). RESULTS: SFAP (40-640 mg/L, 48 h) inhibited the proliferation of SGC-7901 cells, and a positive correlation was noted between inhibitory effects and dosage. At a dosage of 160-320 mg/L in cultured cells, the expression of c-myc was decreased. SFAP (50-200 mg/kg) increased the percentage of CD3 and CD4 T-lymphocytes, the ratio of CD4/CD8, and the contents of Ig such as IgM, IgG or IgA, but decreased the levels of serum sIL-2R in peripheral blood from cyclophosphamide-treated mice. CONCLUSION: SFAP can inhibit the proliferation of SGC-7901 cells via the c-myc gene. In addition, SFAP can modulat the cellular and humoral immunity in cyclophosphamide-induced immunosuppressed mice.     

Lesion of the tuberomammillary nucleus E2-region attenuates postictal seizure protection in rats

Postictal seizure protection (PSP) is an endogenous anticonvulsant phenomenon that follows an epileptic seizure and inhibits the induction of further seizures. The tuberomammillary nucleus (TM), located in the posterior hypothalamus, consists of five subregions and is the sole source of histaminergic neurons in the brain. To determine whether the TM is involved in PSP in rats, we tested the effects of bilateral electrolytic lesions of the TM E2-region on seizures induced by intermittent maximal electroshock (MES). The TM E2-region lesions significantly attenuated PSP during the intermittent MES procedure. Furthermore, intracerebroventricular injection of alpha-fluoromethylhistidine (100 microg), a selective and irreversible histidine decarboxylase inhibitor, mimicked the attenuation of PSP induced by the lesion of TM E2-region. In addition, neurochemical experiments revealed that the TM E2-region lesions markedly decreased basal histamine levels in the cortex, hippocampus, brainstem and hypothalamus, but had no significant effect on basal glutamate and GABA levels. Moreover, intermittent MES induced a persistent decrease of brain histamine levels in both sham-operated and lesioned rats. These results indicate that through its intrinsic histaminergic system, the TM may exert powerful inhibitory function during the intermittent MES procedure and actively participate in the mechanisms of PSP.