Angiographically guided utero-placental gene transfer in rabbits with adenoviruses, plasmid/liposomes and plasmid/polyethyleneimine complexes.

We examined the feasibility of gene transfer to rabbit placenta using adenoviruses, plasmid/liposomes and plasmid/polyethyleneimine (PEI) complexes. Pregnant New Zealand White rabbits (n = 17) were anesthetized and local gene transfer was done via a catheter inserted in uterine arteries under direct angiographic control. Either nuclear targeted LacZ adenoviruses (1.0 x 10(10) p.f.u.), nuclear targeted LacZ plasmid (500 microg)/liposome (DOTMA:DOPE 1:1) complexes or nuclear targeted LacZ plasmid (250 microg)/PEI (25 kDa) complexes (charge ratio +/-4) were used. Animals were killed 3 days later and detection of the transgene expression was done by X-gal staining and RT-PCR. Adenovirus-mediated gene transfer resulted in a high transfection efficiency (34 +/- 10%) in placental trophoplastic cells. Very little, if any, transfection was seen in fetal membranes. Plasmid/liposomes and plasmid/PEI complexes led to a very low (<0.01%) transfection efficiency in trophoblastic cells, but some transfection was seen in fetal membranes. A total of 25 fetuses were analyzed for the presence of transgene at the time of death. In most fetuses expression of the LacZ gene was below the sensitivity of the X-gal staining, but expression was detected by PCR in 50%, 50% and 42% of the analyzed fetuses after adenoviral, plasmid/PEI and plasmid/liposome gene transfer, respectively. No major inflammatory changes were present in the transfected placentas as analyzed by general histology and macrophage- and T cell-specific immunostainings. We conclude that catheter-mediated intravascular gene transfer with adenoviruses can be used for the transfection of placental trophoplastic cells, but plasmid complexes are inefficient for this purpose. However, selective angiographically guided gene transfer also led to leakage of the vector to fetuses. Therefore, if gene therapy is developed for the treatment of placental disorders, the gene-vector combination should not be harmful to the fetus and the expression of the transgene should only occur in placenta.     

荷瘤兔骨髓间充质干细胞在射频消融自体血清诱导下向肝样细胞的分化

目的:鉴于目前对体外培养诱导骨髓间充质干细胞向肝样细胞分化的条件掌握尚不成熟,尤其是无血清培养.实验拟将临床常用微创治疗同细胞移植结合,观察荷瘤兔骨髓间充质干细胞在射频消融兔肝肿瘤自体血清诱导下向肝样细胞的分化,寻求一种自体肝组织的修复方法.方法:实验于2006-09/2007-09在西安交通大学医学院中心实验室完成.实验室为教育部基因与环境研究重点实验室.[1]实验材料:健康新西兰大白兔,二三月龄,雌雄不限,体质量1.5～2.0kg,由西安交通大学医学院动物试验中心提供,实验过程中对动物处置符合动物伦理学标准.荷瘤种兔包块由西安交通大学第一医院介入中心刘亚民教授馈赠.[2]实验方法:取荷瘤种兔瘤组织块,在兔在肝脏右叶做一直径为2~3mm的隧道,将肿瘤组织块植入隧道底部制备肝肿瘤动物模型.射频消融治疗灭活荷瘤兔肝脏肿瘤及边缘1cm正常肝脏组织,于72h后取全血及骨髓.应用体外细胞培养技术自骨髓血中分离、纯化兔骨髓间充质干细胞后,取P2细胞在4种不同培养基培育:对照组:含体积分数为0.1胎牛血清的低糖DMEM培养液;射频兔肝肿瘤治疗血清组:无血清低糖DMEM培养液加入射频兔肝肿瘤治疗血清;射频肝肿瘤治疗血清热灭活组:无血清低糖DMEM培养液加入射频肝肿瘤治疗灭活血清;生长因子组:含0.1胎牛血清的低糖DMEM培养液加入肝细胞生长因子及表皮细胞生长因子.[3]实验评估:倒置显微镜下观查细胞形态变化,流式细胞仪测定细胞周期并观察其增殖及生长特征,免疫荧光法检测肝细胞标志物白蛋白及CK18表达,鉴定细胞性质及功能.结果:[1]原代、传代培养及流式细胞仪结果显示,射频肝肿瘤治疗兔血清及肝细胞生长因子、表皮生长因子对兔骨髓间充质干细胞的诱导能明显促进细胞的增殖,S G2 M期细胞百分数明显增高,2周后免疫荧光染色可见分化细胞表达肝细胞标志物CK18和白蛋白.[2]射频肝肿瘤治疗兔灭活血清及胎牛血清能使兔骨髓间充质干细胞生长,但不如射频肝肿瘤治疗兔血清及肝细胞生长因子、皮细胞生长因子促进细胞增殖明显(P<0.01),传代培养2周,细胞形态未见明显变化,未见肝细胞标志物CK18和白蛋白表达.结论:肿瘤射频消融兔血清可激活、诱导自体骨髓间充质干细胞增殖并向肝样细胞分化,使射频消融肿瘤治疗的同时行肝样细胞移植修复肝损伤成为可能.     

High-pressure liquid chromatographic assay and pharmacokinetics of HR 810 after intramuscular injection in rabbits.

A high-pressure liquid chromatographic assay was developed for the detection of HR 810 in rabbit plasma. There was no interference in the high-pressure liquid chromatographic assay from other antibiotics. The method was accurate, reproducible, and capable of detecting less than 1 microgram of HR 810 per ml in plasma. The assay correlated with the microbiological assay (correlation coefficient, 0.93) and was used to quantitate the concentration of HR 810 in rabbit plasma and determine its half-life subsequent to a 20-mg/kg intramuscular dose. The peak concentration of HR 810 was 51.2 +/- 8.0 micrograms/ml at 1 h postdose. The half-life of the absorption phase (mean +/- standard deviation) was 0.35 +/- 0.10 h, and the half-life of the elimination phase was 0.75 +/- 0.06 h. This is 54% less than the half-life of elimination of 1.38 h previously reported for the intravenous dose.     

55岁及以下乳腺癌术后患者子宫及卵巢病变的监测价值研究

目的：探讨55岁及以下乳腺癌术后患者出现子宫内膜及卵巢病变的临床及超声监测价值 方法：回顾性分析首都医科大学北京妇产医院及首都医科大学大兴教学医院2011年1月至2021年10月以来所有55岁及以下乳腺癌术后患者临床怀疑出现子宫及卵巢病变的病例共148例,所有病例均做出了诊刮或手术取得了子宫内膜织学病理或做出随访观察,根据组织学病理结果应用X2检验及独立样本T检验统计分析出55岁及以下乳腺癌术后患者出现子宫及卵巢恶性病变特点,并应用统计积分量化的方式分析经阴道超声监测子宫内膜恶性病变的价值。 结果：首先根据病例是否应用他莫昔芬TAM治疗分为TAM组87例及非TAM组61例,根据子宫内膜诊刮及手术病理两组出现良性及恶性子宫内膜比例进行统计学比较得出P=0.712;进而对所有子宫内膜良性组共128例及恶性组共20例病例的病人一般资料及超声声像图各指标进行分别统计,两组病例在病人体重指数（P=0.048）、乳腺癌术后时间（P=0.000）、家族史（P=0.005）及是否出现临床症状（P=0.000）上存在明显差异,超声指标上在子宫内膜回声是否均匀（P=0.038）、内膜是否存在囊性变（P=0.038）、内膜有无息肉样变（P=0.000）、有无宫腔积液（P=0.000）及内膜是否存在动脉血流（P=0.000）上均存在明显差异。采用超声指标积分量化的方式评估子宫内膜恶性病变,当积分≥2分时诊断的敏感性及特异性分别为70%及82%,而当超声积分≥3分时诊断的敏感性较低约40%,但特异性较高约96%。本研究148例乳腺癌病人中63例发现卵巢包块,62例为囊性、部分囊内伴有细分隔,1例为囊实性,手术病理均为良性病变或在随访中缩小或消失。 结论：本研究服用TAM的乳腺癌术后患者子宫内膜恶性变的发生率与未服用者比较无明显差异。本研究中子宫内膜恶性病例的总体发生率较高,为13.5%,临床上乳腺癌术后患者出现体重指数增加、乳腺术后时间较长、有恶性肿瘤家族史及出现妇科临床症状的患者出现子宫内膜恶变机率增加;超声图像上显示子宫内膜回声不均、内膜出现息肉样变、宫腔积液及内膜存在动脉血流应警惕出现子宫内膜恶变;而乳腺癌患者出现卵巢病变本组均为良性或可自行吸收。临床上可以尝试应用超声指标积分量化的方式评估子宫内膜恶性病变。     

超声检测骨髓干细胞水凝胶制剂对实验兔心肌梗死后左心室功能恢复的研究

目的 应用超声检测家兔急性心肌梗死(AMI)模型骨髓干细胞水凝胶复合物注射前后心功能,评价该方法治疗心肌梗死的疗效.方法 56只兔建立左室前壁AMI模型,制备BrdU标记骨髓干细胞(BMSCs)水凝胶复合物.AMI后1周存活兔52只分为4组行梗死区心外膜注射:①处理组(A组),BrdU标记BMSCs水凝胶复合物;②阳性对照组(B组),BrdU标记BMSCs;③材料组(C组),BrdU标记后水凝胶;④空白组(D组),等量胎牛血清.分别于AMI造模前、AMI造模后1周及心外膜注射处理后4周对4组兔进行超声检查.常规超声心动图测定左室舒张末期内径(LVDd)、左室前壁舒张末期厚度(AW)、左室射血分数(LVEF),定量组织速度成像(QTVI)分析左室前壁、前室间隔基底段、中段收缩期峰值运动速度(Vs)及舒张早期峰值运动速度(VE).结果 与AMI造模前比较,AMI造模后1周4组AMI模型兔LVDd显著增大,AM厚度显著变薄,LVEF显著减低,梗死局部Vs、VE均显著降低,差异有统计学意义(P＜0.05).与AMI造模后1周比较,心外膜注射后4周材料组与空白组LVDd、AM厚度、LVEF及梗死局部Vs、VE无显著变化;处理组与阳性对照组LVDd显著减小,AM厚度显著增厚,LVEF及梗死局部Vs、VE显著增高,差异有统计学意义(P＜0.05),以处理组变化更显著(P＜0.05).结论 骨髓干细胞水凝胶复合物可有效治疗心肌梗死,改善心肌梗死后左室重构及梗死局部收缩、舒张功能.     

Ultrasound facilitates transduction of naked plasmid DNA into colon carcinoma cells in vitro and in vivo

One approach to improve the efficacy of in vivo gene therapy, with the aim at enhancing expression of the transgene, involves utilization of mechanical forces to facilitate transduction of DNA into cells. In this study, we evaluated the feasibility of mechanical insonation in gene transfers with naked DNA plasmid loading both in vitro and in vivo. We used an ultrasound probe, which can focus the ultrasonic beam in the exit zone of the probe. The reporter pcDNA3-lacZ plasmid, containing Escherichia coli lacZ or the beta-galactosidase gene (beta-gal), and the neomycin 3'-phosphotransferase gene (neo), was used for evaluation of transfer efficiency. Expression of beta-gal in MC38 murine colon carcinoma cells was measured after insonation of 20 W/cm2 with continuous 1.0-MHz wave exposure. In a transient assay, significant numbers of cells were transduced with the beta-galactosidase gene. After cells were treated with geneticin, we also observed a difference in colonogenicity between noninsonated and insonated groups. When MC38 cells were implanted in syngeneic mice and plasmid was injected, the insonation that followed facilitated beta-galactosidase expression. These results indicate that insonation represents a potential approach for gene therapy when combined with naked DNA plasmid injection.     

Uptake, biodistribution, and time course of naked plasmid DNA trafficking after intratumoral in vivo jet injection

Nonviral jet injection is an applicable technology for in vivo gene transfer of naked DNA. However, little is known about the biodistribution and clearance of jet-injected DNA, or about its localization within tissue and cells. Therefore, in this study we analyzed the intratumoral and systemic biodistribution of jet-injected naked DNA in human colon carcinoma-bearing NCr-nu/nu mice, which were jet-injected with the pCMVbeta plasmid DNA. Intratumoral and systemic plasmid DNA biodistribution was analyzed 5, 10, 20, and 40 min and 3, 6, 24, 48, and 72 hr after jet injection, using quantitative real-time polymerase chain reaction. In the tumors, a rapid drop in naked DNA load within 24 hr of jet injection was shown. Detailed analysis of intratumoral distribution of rhodamine-labeled DNA revealed the presence of plasmid DNA within tumor cells 5 min after jet injection and further accumulation of significant DNA amounts in the cell nuclei 30 to 60 min after jet injection. In the blood, DNA amounts rapidly dropped within 10 to 40 min of jet injection to less than 0.001 pg of plasmid per 250 ng of tissue DNA and only minimal plasmid DNA dissemination was detected in liver, lung, spleen, kidney, and ovaries, which was cleared 3 to 6 hr after jet injection. By contrast, in heart, bone marrow, and brain almost no plasmid DNA was detectable.     

Comparison of clinical outcomes and spectral Doppler indices of uterine and ovarian stromal arteries in women undergoing myomectomy with or without hypogastric arterial ligation.

OBJECTIVE: To compare clinical outcomes and hemodynamic alterations of uterine and ovarian stromal arteries between patients with symptomatic myomas undergoing myomectomy preceded by arterial ligation and those undergoing myomectomy alone. METHODS: In this prospective, non-randomized comparative study, myomectomy was performed on 69 women with symptomatic myomas. Myomectomy alone was performed in 31 patients (Group I) and myomectomy with concomitant bilateral hypogastric arterial ligation was performed in 38 patients (Group II). In both groups, surgical results and clinical outcomes were evaluated by peripheral hemoglobin levels, a pictorial blood-loss assessment chart, and visual analog scales. Spectral Doppler indices of uterine and ovarian stromal arteries, including peak systolic velocity, end-diastolic velocity, pulsatility index and resistance index were performed preoperatively, and 1 day and 1 or more months postoperatively. RESULTS: Twenty-two patients in Group I and 31 patients in Group II received regular follow-up examinations for a mean follow-up period of 10.1 months. Menstrual flow, dysmenorrhea and hemoglobin levels improved significantly after surgery in both groups. Blood loss during surgery was less in Group II than it was in Group I (P=0.02). Doppler indices of uterine and ovarian stromal arteries from preoperation to mean follow-up point were not significantly different between the groups, except for a significantly lower uterine artery pulsatility index in Group II (P=0.01). CONCLUSIONS: Myomectomy with hypogastric arterial ligation for symptomatic myomas is as efficient as is myomectomy alone and reduces blood loss during surgery. Serial Doppler studies showed that hypogastric ligation does not block uterine and ovarian perfusion, and even reduces the impedance of the uterine arteries. The long-term recurrence rate after myomectomy with hypogastric arterial ligation remains to be determined.     

红花注射液对实验性脑缺血再灌注损伤中一氧化氮和内皮素的影响

目的　探讨一氧化氮 (NO)和内皮素 (ET)在脑缺血再灌注损伤 (CIRI)中的作用及红花注射液对其的影响。方法　实验家兔分为假手术对照组 (n =8)、脑缺血再灌注组 (n =8)及脑缺血再灌注 +红花组 (n =8) ;分别检测缺血前、缺血 3 0min及再灌注 3 0、60、12 0min 5个时相点血浆NO代谢产物 (NOP)的含量、ET水平的变化 ,同时测定再灌注 12 0min脑组织NOP、ET的浓度 ,并行脑组织电镜观察。结果　脑缺血再灌注 3 0min血浆NOP明显低于假手术对照组 (P <0 0 5 ) ;脑缺血再灌注3 0、60、12 0min血浆ET显著高于假手术对照组 ,尤以再灌注 12 0min变化显著 (P <0 0 1) ;脑组织NOP明显低于、ET显著高于假手术对照组 (P <0 0 5和P <0 0 1) ;脑组织超微结构发生异常改变。红花可逆转上述指标的异常变化。结论　缺血再灌注导致血管内皮功能紊乱 (即NO水平下降和ET水平升高 ) ,在CIRI发生发展中起介导作用 ;红花注射液通过保护血管内皮 ,提高机体内NO水平和降低机体内ET水平 ,从而减轻CIRI。     

Determination of volatile alcohols and acetone in serum by non-polar capillary gas chromatography after direct sample injection

In this method for detection and quantification of volatile alcohols by capillary gas chromatography, the serum sample is deproteinized, then directly injected into the gas chromatograph with 1-propanol as the internal standard. The capillary column is a 30-m bonded methylsilicone-coated, fused-silica column. With helium as the carrier gas, the injector inlet is set at a split ratio of 1/30 and the average linear velocity in the column is 25 cm/s. Injector and flame-ionization detector temperatures are 280 degrees C, oven temperature 35 degrees C. Chromatography time is less than 3 min.     

Longevity of rAAV vector and plasmid DNA in blood after intramuscular injection in nonhuman primates: implications for gene doping

Legitimate uses of gene transfer technology can benefit from sensitive detection methods to determine vector biodistribution in pre-clinical studies and in human clinical trials, and similar methods can detect illegitimate gene transfer to provide sports-governing bodies with the ability to maintain fairness. Real-time PCR assays were developed to detect a performance-enhancing transgene (erythropoietin, EPO) and backbone sequences in the presence of endogenous cellular sequences. In addition to developing real-time PCR assays, the steps involved in DNA extraction, storage and transport were investigated. By real-time PCR, the vector transgene is distinguishable from the genomic DNA sequence because of the absence of introns, and the vector backbone can be identified by heterologous gene expression control elements. After performance of the assays was optimized, cynomolgus macaques received a single dose by intramuscular (IM) injection of plasmid DNA, a recombinant adeno-associated viral vector serotype 1 (rAAV1) or a rAAV8 vector expressing cynomolgus macaque EPO. Macaques received a high plasmid dose intended to achieve a significant, but not life-threatening, increase in hematocrit. rAAV vectors were used at low doses to achieve a small increase in hematocrit and to determine the limit of sensitivity for detecting rAAV sequences by single-step PCR. DNA extracted from white blood cells (WBCs) was tested to determine whether WBCs can be collaterally transfected by plasmid or transduced by rAAV vectors in this context, and can be used as a surrogate marker for gene doping. We demonstrate that IM injection of a conventional plasmid and rAAV vectors results in the presence of DNA that can be detected at high levels in blood before rapid elimination, and that rAAV genomes can persist for several months in WBCs.     

Transfection of CD14 into 70Z/3 cells dramatically enhances the sensitivity to complexes of lipopolysaccharide (LPS) and LPS binding protein

Bacterial endotoxin (lipopolysaccharide [LPS]) causes fatal shock in humans and experimental animals. The shock is mediated by cytokines released by direct LPS stimulation of cells of monocytic origin (monocyte/macrophage [MO]). Recent studies have supported the concept that the plasma protein, LPS binding protein (LBP), plays an important role in controlling MO responses to LPS. Specifically, evidence has been presented to suggest that CD14, a membrane protein present in MO, serves as a receptor for complexes of LPS and the plasma protein LPS binding protein (LBP). In this function CD14 mediates attachment of LPS-bearing particles opsonized with LBP and appears to play an important role in regulating cytokine production induced by complexes of LPS and LBP. The CD14-, murine pre-B cell line 70Z/3 responds to LPS by synthesis of kappa light chains and consequent expression of surface IgM. To better understand the role of CD14 in controlling cellular responses to LPS, we investigated the effect of transfection of CD14 into 70Z/3 cells on LPS responsiveness. We report here that transfection of human or rabbit CD14 cDNA into 70Z/3 cells results in membrane expression of a glycosyl-phosphatidylinositol-anchored CD14. When LPS is complexed with LBP, CD14-bearing 70Z/3 cells bind more LPS than do the parental or 70Z/3 cells transfected with vector only. Remarkably, the expression of CD14 lowers the amount of LPS required to stimulate surface IgM expression by up to 10,000-fold when LPS dose-response curves in the CD14-, parental and CD14-bearing, transfected 70Z/3 cells are compared. In contrast, the response of CD14-bearing 70Z/3 cells and the parental 70Z/3 cell line (CD14-) to interferon gamma is indistinguishable. LPS stimulation of the parental and CD14-bearing 70Z/3 cells results in activation of NF-kB. These data provide evidence to support the concept that the LPS receptor in cells that constitutively express CD14 may be a multiprotein complex containing CD14 and membrane protein(s) common to a diverse group of LPS-responsive cells.     

变异链球菌表面蛋白SpaP P区真核表达质粒的构建及表达

背景:变异链球菌是龋病的主要致龋菌,针对介导变异链球菌非蔗糖依赖性黏附的重要毒力因子SpaP的基因疫苗在理论上能对抗变异链球菌黏附于牙面和进一步破坏牙体硬组织.目的:构建变异链球菌表面蛋白P区(SpaP/P)真核表达质粒pVAX1-spapiP,并观察其在哺乳动物细胞COS.7中的表达.方法:通过基因重组技术,构建真核表达质粒pVAX1-spap/P,并经酶切分析、测序分析鉴定正确后,采用脂质体转染法,将其转染至COS-7细胞中,然后经免疫组织化学SABC法检测其在细胞中的表达.结果与结论:真核表达质粒pVAX1-spap/P经EcoR Ⅰ和Xba Ⅰ双酶切分析,证实携带1.2 kb的目的基因spapiP片段,经测序分析,目的基因正向插入到预先设计的载体位点处.pVAX1-spap/P转染的细胞胞质呈褐色,pVAX1空载体质粒转染的细胞胞质中无着色.证实实验成功构建真核表达质粒pVAX1-spap/P,所携带的基因序列正确,能够在真核细胞COS-7中正确表达目的蛋白.     

Pharmacokinetics of sulbactam/ampicillin in humans after intravenous and intramuscular injection

We investigated the pharmacokinetic properties of sulbactam/ampicillin (S/A), after intravenous (0.5/1.0 and 1.0/2.0 g) and intramuscular (0.5/1.0 g) coadministration in 10 male subjects. After 1.0/2.0 g intravenous doses of S/A the half-lives (t1/2 beta) were 1.14 +/- 0.14/1.09 +/- 0.16 h. The values for plasma clearance (CLp) were 198.83 +/- 26.27/250.33 +/- 39.28 ml/min and the renal clearance (Clr) 173.50 +/- 19.66/208.80 +/- 26.43 ml/min. The post distributive volumes (V beta) were 19.67 +/- 3.24/23.56 +/- 5.76 liters. Similar values were obtained after 0.5/1.0 g of S/A intravenous coinjection. After 0.5/1.0 g intramuscular coadministration the t1/2 beta values were 1.26 +/- 0.18/1.20 +/- 0.15 h. The values for Clp were 208.00 +/- 28.73/243.17 +/- 33.24 ml/min, for Clr 179.50 +/- 20.26/202.67 +/- 27.61 ml/min and for V beta 22.27 +/- 4.12/25.30 +/- 4.87 liters. The renal clearance of sulbactam is comparable to that of ampicillin and both clearances are greater than the glomerular filtration rate, suggesting active renal tubular secretion of the two drugs. The large volumes of distribution, and the ratio K12/K21 = 0.5 show the extensive distribution of the two drugs into extracellular fluids. The very similar values of the pharmacokinetic parameters of sulbactam and ampicillin confirm that the kinetics of the two drugs closely resemble one another.     

Dendritic cells matured with TNF can be further activated in vitro and after subcutaneous injection in vivo which converts their tolerogenicity into immunogenicity

Dendritic cell (DC) maturation can occur by different types of stimuli. Previously, we described that murine DC matured with tumor necrosis factor (TNF) up-regulate surface MHC and costimulatory molecules but lack cytokine release, and therefore termed them semi-mature DC. These TNF/DC-induced tolerance after intravenous (i.v.) injection in a model of experimental autoimmune encephalomyelitis (EAE). Here, we show that TNF/DC are not terminally differentiated but can still respond to the microbial stimulus lipopolysaccharide. Subcutaneously injected TNF/DC induce an unpolarized T(H)1/T(H)2 response and are not protective in the experimental autoimmune encephalomyelitis model. Although TNF/DC home to the draining lymph node, they remain negative for intracellular cytokine stainings. However, the nonmigrating, endogenous DC started to produce interleukin (IL)-12p40, TNF and little IL-6, IL-10, and MCP-1 in a bystander fashion. Together, DC matured with the inflammatory stimulus TNF remains responsive to further signals in vitro and in vivo. These signals can be provided by pathogens or the subcutaneous injection route, which can convert them from tolerogenic to immunogenic DC. These findings are important for selecting the appropriate injection route of human DC for tumor immunotherapy.     

Serum markers of monocyte/macrophage activation in patients with Alzheimer's disease and other types of dementia

OBJECTIVES: Recently an increase in serum neopterin has been described in patients with Alzheimer's disease (AD) that would be associated with an increased cell-mediated immune response. We have studied the serum levels of several monocyte/macrophage activation markers in patients with AD and other types of dementia. DESIGN AND METHODS: Serum neopterin concentration, and the chitotriosidase (ChT), angiotensin-converting enzyme (ACE) and adenosine deaminase (ADA) activities were determined in 30 patients with AD, in 19 patients with other types of dementia, and in 24 nonaffected controls. RESULTS: Neopterin concentration was significantly higher in the subgroup of AD patients with a global deterioration scale higher than in the other patients with AD, patients with other types of dementia and in the control group (p < 0.005). However, the activities of ChT, ACE and ADA, despite having a significant correlation with neopterin, did not present any statistically significant differences among the groups studied. CONCLUSION: In the most advanced clinical stages of AD, as well as an increased immune activation, an impaired formation of tetrahydrobiopterin from dehydroneopterin triphosphate would contribute to an increase in the serum concentration of neopterin. However, the large overlap between the groups, limits the possible clinical value of serum neopterin in AD patients.     

股动脉周围骨骼肌多点注射携带肝细胞生长因子基因重组质粒糖尿病大鼠肢体缺血骨骼肌的组织学变化

背景:质粒载体因其较好的生物安全性和较高的体内维持时间,被广泛应用于基因治疗研究领域。目的:观察携带肝细胞生长因子基因重组质粒pUDKH在糖尿病后肢缺血模型大鼠股动脉周围骨骼肌多点注射15d后骨骼肌组织的病理学变化。方法:利用STZ制备SD大鼠糖尿病模型。随机分为3组,建模后24h内高浓度组注射pUDKH 200μg/只,低浓度组注射pUDKH 100μg/只,对照组注射等体积医用注射用水。15d后,取大鼠的骨骼肌组织进行病理学观察。结果与结论:对照组肌浆萎缩变性,纤维化增生并透明变性,浆细胞浸润变性,肌间隙变宽,有局灶性脓肿形成;pUDKH治疗后有丰富的微血管形成,高浓度组肌纤维横纹较低浓度组保持完整。提示携载肝细胞生长因子基因质粒pUDKH对糖尿病大鼠肢端缺血有治疗作用。     

斑点追踪成像评价实验兔心肌梗死后骨髓干细胞移植区域心肌收缩功能

目的 应用超声斑点追踪(STI)技术探讨兔左室心肌梗死区域骨髓干细胞移植后的心肌收缩功能.方法 24只健康新西兰兔,随机分为3组:正常对照组(假开胸组)、急性心肌梗死(AMI)组(结扎左冠状动脉前降支,造成左室前壁心肌梗死)和骨髓干细胞移植组(AMI后2周,对梗死区域进行干细胞移植),于骨髓干细胞移植后4周分别采集三组动物左室短轴基底段、中间段及心尖段的动态二维灰阶图像;应用STI技术分析左室不同水平各节段心内膜下心肌径向应变率(SrR)、周向应变率(SrC)、旋转率(RotR)及扭转角度(Rot),常规超声心动图测定左室舒张末期内径(LVEDd)、左室射血分数(LVEF)及左室短轴缩短率(LVFS),心导管测量左室收缩压(LVSP)、左室舒张末期压(LVEDP)、左室压力最大上升及下降速率(LVdp/dtmax).结果 与正常对照组比较,AMI组LVEDd显著增大,LVEF、LVFS显著减低,LVSP、LVdp/dtmax明显降低,LVEDP明显升高,左室前壁局部及左室短轴三水平整体SrR、SrC、RotR和左室Rot均降低;骨髓干细胞移植组LVEDd较AMI组显著减小,LVEF、LVFS明显升高,LVSP、LVdp/dtmax明显升高,LVEDP降低明显,左室前壁局部及左室短轴三水平整体SrR、SrC、RotR和左室Rot均升高,但与正常对照组比较差异无统计学意义.结论 STI可准确评价骨髓干细胞移植区域的心肌功能.

Abstract：

Objective To assess the left ventricular(LV) regional myocardial systolic function after bone marrow mesenchymal stem cells (BMSCs) injection in rabbits with acute myocardial infarction(AMI) by 2-dimensional ultrasound speckle-tracking imaging.Methods Twenty-four healthy rabbits were randomly divided into three groups:group of sham-operated,group of masculine control (AMI was induced by ligation of left anterior descending coronary artery),and group of cell infusion(after two weeks of AMI,bone marrow mesenchymal stem cells were injected to the region of AMI).Four weeks after cell deliver two-dimensional strain images were acquired from LV short-axis view(at the levels of mitral annulus,muscle papillary and apex),radial strain rate (SrR),circumference strain rate (SrC),rotation rate (RotR) and rotation (Rot) of three levels in short-axis views were measured by speckle-tracking imaging.The conventional echocardiography indices included LV diameter of end diastole(LVEDd),LV ejection fraction(LVEF),LV fractional shortening(LVFS).The catheter indices included LV systolic pressure(LVSP),LV end diastolic pressure (LVEDP)and maximum rate of rise and descend of LV pressure(LVdp/dtmax).ResultsCompared with sham-operated group,rabbits had significantly larger LVEDd,lower LVEF and LVFS,lower LVSP and LVdp/dtmax,higher LVEDP in control group,SrR,SrC,RotR and Rot of LV anterior regional myocardial function and global myocardial function of three levels in short-axis views were lower.Compared with control group,the rabbits had significantly smaller LVEDd,larger LVEF and LVFS,larger LVSP and LVdp/dtmax,lower LVEDP,SrR,SrC,RotR and Rot of LV anterior regional myocardial function and global myocardial function of three levels in short-axis views were larger in group of cell infusion.There was no significant between group of cell infusion and group of sham-operated.Conclusions Speckle tracking imaging can evaluate the regional myocardial systolic function of the area of BMSCs transplantation accurately.     

Sustained expansion of NKT cells and antigen-specific T cells after injection of alpha-galactosyl-ceramide loaded mature dendritic cells in cancer patients

Natural killer T (NKT) cells are distinct glycolipid reactive innate lymphocytes that are implicated in the resistance to pathogens and tumors. Earlier attempts to mobilize NKT cells, specifically, in vivo in humans met with limited success. Here, we evaluated intravenous injection of monocyte-derived mature DCs that were loaded with a synthetic NKT cell ligand, alpha-galactosyl-ceramide (alpha-GalCer; KRN-7000) in five patients who had advanced cancer. Injection of alpha-GalCer-pulsed, but not unpulsed, dendritic cells (DCs) led to >100-fold expansion of several subsets of NKT cells in all patients; these could be detected for up to 6 mo after vaccination. NKT activation was associated with an increase in serum levels of interleukin-12 p40 and IFN-gamma inducible protein-10. In addition, there was an increase in memory CD8+ T cells specific for cytomegalovirus in vivo in response to alpha-GalCer-loaded DCs, but not unpulsed DCs. These data demonstrate the feasibility of sustained expansion of NKT cells in vivo in humans, including patients who have advanced cancer, and suggest that NKT activation might help to boost adaptive T cell immunity in vivo.