Limbal Approach-Subretinal Injection of Viral Vectors for Gene Therapy in Mice Retinal Pigment Epithelium

The eye is a small and enclosed organ which makes it an ideal target for gene therapy. Recently various strategies have been applied to gene therapy in retinopathies using non-viral and viral gene delivery to the retina and retinal pigment epithelium (RPE). Subretinal injection is the best approach to deliver viral vectors directly to RPE cells. Before the clinical trial of a gene therapy, it is inevitable to validate the efficacy of the therapy in animal models of various retinopathies. Thus, subretinal injection in mice becomes a fundamental technique for an ocular gene therapy. In this protocol, we provide the easy and replicable technique for subretinal injection of viral vectors to experimental mice. This technique is modified from the intravitreal injection, which is widely used technique in ophthalmology clinics. The representative results of RPE/choroid/scleral complex flat-mount will help to understand the efficacy of this technique and adjust the volume and titer of viral vectors for the extent of gene transduction.     

丹红注射液对急性冠脉综合征患者血浆脑钠肽及心率变异性的影响

目的:探讨丹红注射液对急性冠脉综合征患者血浆脑钠肽水平及心率变异性的影响。方法:选取我院收治的急性冠脉综合征患者72例,并将其随机分为实验组和对照组。对照组36例,予以阿司匹林肠溶片100 mg/日一次口服、低分子肝素钠200 IU/kg体重,每日一次皮下注射及普萘洛尔20 mg/日3次口服等常规治疗,2周为一个疗程,治疗两个疗程;实验组36例,在常规治疗的基础上加用丹红注射液,30 m L,加入5%葡萄糖注射液250 m L滴注,每日二次,2周为一个疗程,治疗两个疗程。治疗结束后,比较治疗前后两组患者的血浆脑钠肽水平及心率变异性中SDNN、SDANN、RMSSD、PNN50%变化。结果:1治疗后,两组患者血浆脑钠肽水平均有所改善,且实验组血浆脑钠肽(96.72±19.43 ng/L)较对照组血浆脑钠肽(147.53±26.89 ng/L)明显下降,差异有统计学意义(P0.05);2治疗后,两组患者心率变异性中SDNN、SDANN、RMSSD、PNN50%各指标均有所改善,且实验组改善情况明显优于对照组,差异有统计学意义(P0.05)。结论:丹红注射液能够明显降低急性冠脉综合征患者的血浆脑钠肽水平,并显著改善心率变异性各项参数,从而可能有助于改善记性冠脉综合征患者的预后。     

丹参酮IIA磺酸钠注射液辅助治疗对突发性耳聋患者血液流变学及临床疗效的影响

目的：探讨丹参酮IIA 磺酸钠注射液辅助治疗对突发性耳聋患者血液流变学及临床疗效的影响。方法：选取我院收治的突发 性耳聋患者78 例,并将其随机分为实验组和对照组,每组39 例。对照组给予三磷酸腺苷二钠、辅酶A、复合维生素、双密达莫片 及高压氧治疗,而实验组在对照组的基础上给予丹参酮IIA 磺酸钠注射液治复合组治疗。观察和比较两组患者治疗后的听力恢复 等级分布以及治疗前后的电测听情况和各项血液流变学指标的变化情况。结果：治疗后,与对照组比较,实验组听力恢复Ⅰ级患 者的比例更高,电听力平均值均显著降低,各项血液流变学指标均明显改善,差异均有统计学意义(P<0.05)。结论：丹参酮IIA 磺酸 钠注射液辅助治疗能够显著提高突发性耳聋的临床疗效,这可能其改善患者的血液流变学有关     

黄芪注射液对慢性心力衰竭患者BNP水平的影响

目的:探究黄芪注射液对慢性心力衰竭患者外周血清B型脑利钠肽水平的影响。方法:选择2013年12月-2015年1月我院收治的慢性心力衰竭住院患者334例,按用药不同分为实验组和对照组。对照组患者采取常规药物口服治疗,实验组患者应用黄芪注射液静脉滴注。观察并比较两组患者治疗前后外周血B型脑利钠肽水平的变化情况。结果:治疗后,两组患者外周血B型脑利钠肽水平均明显下降,且实验组下降更明显,实验组Ⅰ级、Ⅱ级患者比例较对照组显著升高,差异均具有统计学意义(P0.05)。结论:黄芪注射液能够降低慢性心力衰竭患者的外周血B型脑利钠肽水平,改善其心功能,值得临床推广应用。     

醒脑静注射液对急性脑出血患者FIB和D-二聚体水平的影响

目的:研究醒脑静注射液对急性脑出血患者血浆纤维蛋白原和D-二聚体水平的影响。方法:选取我院神经内科收治的急性脑出血患者90例,随机分为两组,其中对照组45例,予降低颅内压、控制血压、抗感染、营养脑细胞等常规治疗。实验组45例,在常规治疗的基础上加用醒脑静注射液。对比治疗前后患者血浆纤维蛋白原(FIB)、D-二聚体(D1-D)的含量及临床疗效。结果:治疗后,两组患者病情均有所改善,实验组有效率(91.11%)与对照组(82.22%)比较明显较高,差异有统计学意义(P0.05);两组患者血浆FIB含量较治疗前均明显升高,但实验组高于对照组,差异有统计学意义(P0.05);实验组血浆D-D含量明显降低,对照组明显升高,实验组明显低于对照组,差异有统计学意义(P0.05)。结论:醒脑静注射液能够使急性脑出血患者血浆纤维蛋白原含量增加,D-二聚体水平下降,具有良好的临床疗效,值得进一步研究推广。     

Strike-induced chemosensory searching in venomoid pit vipers at Hogle Zoo

Six pit vipers with ligated and severed venom ducts were each observed twice. In the no-strike trial a rodent was presented for 3 sec but held (on tongs) out of striking range. The strike trial began with a 3 sec presentation, but the prey item was then moved into striking range, and all snakes immediately struck and released the rodents. Prey were removed after no-strike and strike presentations and the rate of tongue flicking was recorded for 30 min. Only strike presentations were followed by high rates of tongue flicking, which were indistinguishable from those seen in pit vipers with functioning venom apparatus. We conclude that venom injection is irrelevant for the causation of strike-induced chemosensory searching.     

Inhibition of the system luminol-H2O2-Fe(CN)63− chemiluminescence by the Mn(II) indirect determination of isoniazid in a pharmaceutical formulation

A flow injection procedure for the indirect chemiluminescent determination of isoniazid is proposed. The method is performed in a flow-injection manifold provided with a solid-phase reactor. The reactor was made from manganese dioxide physically entrapped by polymerization; the redox reaction isoniazid–manganese dioxide released Mn(II) which was monitored through its inhibitory effect on the reaction between luminol and hydrogen peroxide in presence of potassium hexacyanoferrate(III). The procedure resulted in a linear calibration graph over the range 5–15 mg/L of isoniazid with a sample throughput of 43 samples/h. The influence of foreign compounds was studied and the method was applied to determination of the drug in a pharmaceutical formulation. © 1998 John Wiley & Sons, Ltd.     

Fertilization of porcine oocytes following intracytoplasmic spermatozoon or isolated sperm head injection

We demonstrated normal fertilization processes (as determined by pronuclear formation, pronuclear apposition and syngamy) in porcine oocytes either following intracytoplasmic spermatozoon (ICSI) or isolated sperm head injection. Microtubule organization and chromatin configuration were investigated in these oocytes during the first cell cycle. Following ICSI, the microtubular aster was organized from the neck of the spermatozoon and filled the whole cytoplasm. These male-derived microtubules appear to move both pronuclei to the center of oocytes. These cytoskeletal changes are analogous to those seen following conventional fertilization. In contrast, following isolated sperm head injection, the sperm aster was not seen. Instead, the microtubule matrix was organized from the cortex and then filled the whole cytoplasm in all cases in normally fertilized oocytes following injection (n = 35). This organization is similar to what has been shown in the parthenogenetically activated oocytes. Chromosome analysis revealed that the oocytes injected with isolated sperm heads were fertilized normally. At 7 days following injection, the incidence of blastocoele formation following ICSI (38%) and isolated sperm head injection (22%) was higher than that following sham injection (2%). These results suggested that successful fertilization and preimplantation development occurred in porcine oocytes following either ICSI or isolated sperm head injection. Our results also indicated that fertilization processes can occur by self-assembled microtubules within cytoplasm in the absence of a sperm centrosome. Mol. Reprod. Dev. 51:436–444, 1998. © 1998 Wiley-Liss, Inc.