Objective To introduce an improved extraction method of prefrontal cortical and striatal synaptosomes from SHR rat. Methods Synaptosomes were prepared from SHR rat brain tissue by Percoll density gradient centrifugation.Transmission electron microscopy was used to assess the morphology and structural integrity of the synaptosomes.Results The obtained synaptosomes showed oval structures surrounded by an intact membrane.Presynaptic components contained one or more mitochondria and a large number of synaptic vesicles.The synaptic clefts were clearly visible, and prominent part of the characteristic compact structure was clear, complete and with higher electron-density. The synaptosome presynaptic membrane, synaptic cleft, and postsynaptic membrane were well preserved, and the synaptosomes were densely distributed, showing typical morphological characteristics of synaptosomes.Conclusions The results of our study improved the traditional preparation method and provide a less time-consuming, highly productive protocol for preparation of structurally typical and intact synaptosomes, suitable for further research on neuroscience and neurological diseases.

Objective To discuss the research status of TCM therapy for ADHD in China in recent ten years; To provide references for clinical workers. Methods Clinical research papers about TCM therapy for ADHD in CNKI, VIP, and CBM from 1st January 2005 to 1st April 2015 were retrieved. Metrology analysis was conducted from the aspects of publication time, diagnostic criteria, contrasted medicine, and types and features of TCM therapy. Results By eliminating duplication literature, 670 papers were screened out. After the screening of exclusion criteria, 148 papers were included. DSM-Ⅳ and CCMD-3 were the most commonly used diagnostic criteria in the 148 papers; internal therapy was the main therapy in TCM therapy; decoction was the main dosage form; liver-kidney yin deficiency syndrome and heart-liver fire syndrome were the main patterns of syndrome; Acori Tatarinowii Rhizoma, Rehmanniae Radix Praeparata and other 19 Chinese medicinal herbs were the main Chinese mateia medica; Xiaoer Zhili Syrup was the most commonly used Chinese patent medicine; acupuncture and moxibustion and auricular point therapy were the main external therapy, and external combined with internal therapy had the best efficacy. Conclusion TCM therapy for ADHD is with high effectiveness. External combined with internal therapy has the best efficacy.

Objective To investigate the neuroprotective effect of acupuncture combined with a Chinese medicine, Danhong injection, on focal cerebral ischemia/reperfusion injury in rats.Methods Middle cerebral artery ( MCA ) occlusion was prepared by putting a nylon suture in the MCA.The MCA blood flow in the animals was restored at two hours after the MCA occlusion by withdrawing the suture to the external carotid artery (ECA).The acupoints“Baihui”(DU20) and “Zusanli”(ST36) were acupunctured and stimulated by twirling the acupuncture needle after reperfusion, and the rats were also administered with Danhong injection by intravenous injection, once every day for three days.Zea-Longa scores were recorded to assess the changes of neurological function. The rats were sacrificed at 72 h after reperfusion. Triphenyltetrazolium chloride ( TTC) staining was used to measure cerebral infarction size.Western blot was used to test the level of Bcl-2 and Bax in the ischemic penumbra.Results The acupuncture combined with Danhong injection treatment group displayed markedly lower neurologic deficit scores, reduced ischemic infarct surface, increased expression of Bcl-2 protein, and reduced expression of Bax protein, compared with those in the model group and Danhong injection alone group.Conclusions Acupuncture combined with Danhong injection show more significant neuroprotective effect than Danhong injection treatment alone on rat models of cerebral ischemia/reperfusion injury.

Objective:To investigate the effects of Maijun'an Tablets on expression of endothelin-1(ET-1) and nitricoxide synthase(NOS) in tissues and plasma of spontaneous hypertensive rats (SHR). Methods:12 spontaneous hypertensive rats (SHR) were divided randomly into normal control group and drug administration group. Rats in drug administration group were orally given with Maijun'an Tablets (hydrochlorothiazide 10 mg?kg-1.d-1,puerarin 36 mg?kg-1.d-1 and total alkaloid of rhynchophylla 122 mg?kg-1.d-1) once a day for 4 weeks,and rats in control group were orally dosed with equal amount of physiological saline. All animals were sacrificed after 4 weeks. The mRNA and protein contents of ET-1 and NOS in plasma,liver,kidnay and blood vessel were determined. Results:Compared with the control group,Maijun'an Tablets can effectively reduce blood pressure of SHRs,regulate down the content of ET-1 and regulate up the content of NOS at mRNA and protein levels. Conclusion:The up-regulation of NOS and down-regulation of ET-1 at mRNA and protein levels were the major antihypertensive machnisam of Maijun'an Tablets for SHRs.

Objective To establish a Real-time quantitative PCR method (qPCR) for the detection of diatom UPA barcoding genes and evaluate its application in the drowning diagnosis. Methods The homologous sequences of diatoms UPA gene was obtained by Blast from GeneBank, based on which the universal primers for diatoms were designed. DNA were extracted from 2 common human symbiotic bacteria (Escherichia coli, Bifidobacterium longum), 3 species of planktonic bacteria, 15 species of planktonic algae, tissue samples (lung, liver and kidney) from human cadavers (28 drowning victims, 1 victims by non-drowning in the water, 3 victims deaths on land) in 32 cases. The specificity, sensitivity and repeatability of the designed primers were tested. The positive rates of diatoms detection in the drowning cases were calculated. The results of the real-time quantitative method were evaluated comparatively by Microwave Digestion-Vacuum Filtration-Automated Scanning Electron Microscopy (MD-VF-Auto SEM) and PCR-Capillary Electrophoresis (PCR-CE). Results The results showed that the primers UPA99 had strong specificity for the diatomaceae (Synedra radians, Navicula sp., Melosira varians, Cyclotella sp. and Nitzschia sp.) DNA. The melting curve of the amplified product was smooth; the peak was narrow; the melting temperature was (87±1)℃. The sensitivity of qPCR method was 1.56×10-5ng/μL with the detection range of 1.56×102ng/mL~1.56×10-5ng/μL, in contrast with the PCR-CE method (1.56×10-3ng/μL). This real-time PCR method showed high repeatability and stability with the coefficient of variation less than 2%. The detection rate of lung, liver and kidney was 89.3%, 71.4% and 64.3% respectively. Conclusion The established qPCR method, based on the universal primers designed for diatom UPA gene, has high specificity, high sensitivity and good repeatability. With a promising prospect for application, qPCR is suitable for drowning diagnosis.

Alzheimer's disease (AD), the most common neurodegenerative disorder, is characterized by memory loss and cognitive dysfunction. The accumulation of misfolded protein aggregates including amyloid beta (Aβ) peptides and microtubule associated protein tau (MAPT/tau) in neuronal cells are hallmarks of AD. So far, the exact underlying mechanisms for the aetiologies of AD have not been fully understood and the effective treatment for AD is limited. Autophagy is an evolutionarily conserved cellular catabolic process by which damaged cellular organelles and protein aggregates are degraded via lysosomes. Recently, there is accumulating evidence linking the impairment of the autophagy-lysosomal pathway with AD pathogenesis. Interestingly, the enhancement of autophagy to remove protein aggregates has been proposed as a promising therapeutic strategy for AD. Here, we first summarize the recent genetic, pathological and experimental studies regarding the impairment of the autophagy-lysosomal pathway in AD. We then describe the interplay between the autophagy-lysosomal pathway and two pathological proteins, Aβ and MAPT/tau, in AD. Finally, we discuss potential therapeutic strategies and small molecules that target the autophagy-lysosomal pathway for AD treatment both in animal models and in clinical trials. Overall, this article highlights the pivotal functions of the autophagy-lysosomal pathway in AD pathogenesis and potential druggable targets in the autophagy-lysosomal pathway for AD treatment.