In order to identify Peucedani Radix, Peucedani Decursivi Radix and their adulterants, the internal transcribed spacer 2 (ITS2) regions of Peucedani Radix, Peucedani Decursivi Radix and their adulterants were amplified and bidirectionally sequenced based on the Principles for Molecular Identification of Traditional Chinese Materia Medica Using DNA Barcoding, which has been promulgated by Chinese Pharmacopoeia Commission. Sequences were analyzed and assembled by Codon Code Aligner V3. 7.1. The relevant data were analyzed by MEGA 5. 0. Species identification analyses were performed by using the nearest distance methods and neighbor-joining (NJ) methods. The result showed that the ITS2 sequence lengths of Peucedani Radix were 229-230 bp and the average intra-specific genetic distances were 0.005. The ITS2 sequence lengths of Peucedani Decursivi Radix were 227 bp and the sequences contained no variation site. The average inter-specific K2P genetic distance of Peucedani Radix, Peucedani Decursivi Radix and their adulterants species were 0.044 and 0.065 respectively. The minimum inter-specific divergence is larger than the maximum intra-specific divergence of Peucedani Decursivi Radix. The nearest distance methods and NJ trees results indicated that Peucedani Radix, Peucedani Decursivi Radix and their adulterants species could be identification clearly. The ITS2 regions can stably and accurately distinguish Peucedani Radix, Peucedani Decursivi Radix and their adulterants.
Apiaceae ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Ribosomal Spacer ; Drug Contamination

Objective To establish a method for detecting HBV cccDNA in hepatocytes of chronic hepatitis B patients.Method 21 liver biopsies from the hepatic operation patients in the hospital of jiangsu province,concluding 19 HBV chronic infected patients (10 HBeAg positive patients and 9 HBeAg negative patients) and 4 uninfected patients,HBV DNA(+) serum of hepatitis B patients was thought as rcDNA.To use proteinase K to release HBV cccDNA and genomic DNA,then divide the cell lysis solution into two parts,one for detecting HBV cccDNA,the other for detecting the number of ?-Globin as internal control. Nucleic acid for detecting HBV cccDNA extracted by phenol-chloroform was digested by plasmid-safe ATP dependent DNase which was applied to digest the single strand DNA in rcDNA and ssDNA,then was quantitated by the primers spanning across the nick and SYBR Green Ⅰ dye.The specifity of PCR production was confirmed by the sequence analysis and rcDNA comparison.The significance of the difference of HBV cccDNA level between HBeAg(+) and HBeAg(-) group was analyzed by two group t test.Results The agarose gelelectrophoresis showed the molecular weight of the PCR production was about 350bp.The coincidence rate of PCR production and goal fragement was nearly 99% by sequence analysis.The result of PCR detection of rcDNA group was negative.The positive rate of HBV cccDNA of liver biopsies of HBeAg (+) patients detected by this method was 100%,the level of HBV cccDNA in the liver biopsies of HBeAg (+) patients was higher than HBeAb(+) patients.Conclusions The specificity of the method is proved by agarose electrophoresis,gene sequencing of the PCR product and rcDNA comparison.The quantitative method that use SYBR Green Ⅰ dye and ?-Globin as internal control is more specific,sensitive and economical,and more suitable for clinical purpose.

Objective To observe the diurnal variation of intraocular pressure(IOP)measured with Goldmann applanation tonomoter(GAT)in patients with primary open angle glaucoma(POAG),normal tension glaucoma(NTG)and healthy controls,and to compare the differences of diurnal variation curves between two eyes of each subject and define the distribution of the peak of IOP. Design Prospective case series.Participants POAG patients,NTG patients and healthy controls.Methods The diurnal variation of IOP in the two eyes of each of 30 POAG patients,30 NTG patients and 30 normal controls were investigated and the distributions of the peak IOP were observed.Main Outcome Measure Intraocular pressure.Results Measured with GAT,6.7% right eyes and 10.0% left eyes of normal people,20.0% right eyes and 23.3% left eyes of NTG patients,and 23.3% right eyes and 20.0% left eyes of POAG patients showed peak IOP at the time points of out-office period.Conclusions POAG patients,NTG patients and normal controls have asymmetric IOP diurnal variation.The two eyes of one individual cannot always be treated as the same.The IOP peak can occur at the time point during the out-office time.Measure IOP in office-time only may miss the dynamic information of IOP.

Objective To investigate the effects of heat-killed Staphylococcus aureus (HKSA) on the apoptosis and expression of iNOS and IL-1β in THP-1 monocytes in the presence of high concentration of glucose.Methods THP-1 cells were cultured in medium containing 25.0 mmol/L(HG) or 5.5 mmol/L (LG,control) D-glucose for 12 h-8 d.The THP-1 cells cultured for 6 d were extracted on the 0-48 h with or without HKSA,then apoptosis and expression of iNOS and IL-1β were examined.Apoptosis was analyzed by flow cytometry and expressions of IL-1β and iNOS were quantitated by real-time PCR.Results The expression of iNOS and IL-1β in THP-1 monocytes was increased significantly in the presence of high concentration of glucose for 12-48 h(P＜0.05),reaching the highest level at 24 h and returned to baseline after 4 d.The expression was significantly lower than that of control after 4-6 d.Apoptosis rate was also increased significantly after 48 h to 4 days.HKSA infection enhanced apoptosis,but inhibited the expression of iNOS and IL-1 β in the presence of high concentration of glucose.The expression of iNOS and IL-1β increased significantly at 6 h(P＜0.01),reaching the highest level at 12 h,but the levels were significantly lower than those in control groups (P＜0.05).Conclusion These data suggest that high concentration of glucose can interfere with the anti-bacterial function of monocytes by reducing their expression of iNOS and IL-1β and enhancing their apoptosis.

Objective To explore the prevalence of visual,hearing,speech,physical,intellectual and mental disabilities in China,2006.Methods The reference time of the Second National Sample Survey on Disability was zero hour,April 1,2006,and 2 526 145 individuals were investigated from 31 provinces autonomous regions or municipalities under the"Disability Classification and Standards for the Second National Sampling Survey on Disability".The classified prevalence rates of 6 types of disability was analyzed.Results (1)The overall prevalence of disabilities was 6.39%,which was 1.49 percent higher than the prevalence rate in 1987.The prevalence of physical disability was the highest(2.34%),while the speech disability was the lowest(0.53%).(2)29.49% of all the disabled persons were classified as severe who suffered grade-1 and grade-2 while 70.51% of them were moderately and mildly disabled suffering from grade-3 and grade-4.Over 65% of the speech disability and over 45% of the mental disability were identified.which were much higher than the other 4 types of disability.(3)The correlation between age,gender.residential place and each of 6 types of disability were statistically significant(P＜0.001).(4)Among all the disabling factors of speech disability and mental retardation disability,congenital factors made great contribution(22.67% and 22.41% respectively).Conclusion According to the analysis results,the characteristics of disabled persons in China were as follows:ageing population had high prevalence rate of disability;all the population mainly suffered moderate and mild disability,but the proportion of severe disability was not low;the prevalence rate was higher and the main reasons caused disability were diversification in rural area.

To introduce the application of Family-Based Association Test (FBAT) in detecting the association or linkage between marker alleles and phenotype or traits in genetic epidemiology. A brief overview on basic steps involved in design, implementation, principles of application, available software as well as the use and feature of FBAT method, were explained based on data from nuclear family. Advantages of FBAT were compared to conventional genetic statistics approaches. Data showed that this approach might make the most use of parental genotype,genotype of affected offspring and their phenotype to test the association between biallelic or multi-allelic markers, phenotype or traits to their conditional distributions given the minimal sufficient statistics under the null hypothesis for the genetic model. Covariates, such as traits or environmental exposures, might be included into this proudure so as to effectively control the bias of population admixture through adjustment of variables. FBAT could be used for genetic analysis with data from nuclear family since it is more advantageous than the conventional statistical methods.

DNA barcoding is a rapidly developing frontier technology in the world and will be useful in promoting the quality control and standardization of traditional Chinese medicine. Until now, many studies concerning DNA barcoding have focused on leaf samples but rarely on Chinese herbal medicine. There are three issues involved in DNA barcoding for traditional Chinese medicinal materials: (1) the extraction methods for total DNA of the rhizomes of the medicinal materials; (2) intra-specific variation among samples from different places of origin; (3) accuracy and stability of this method. In this study, Gentianae Macrophyllae Radix was used to verify the stability and accuracy of DNA barcoding technology. Five regions (ITS2, psbA-trnH, matK, rbcL, and ITS) were tested for their ability to identify 86 samples of Gentianae Macrophyllae Radix and their adulterants. After improving the DNA extraction method, genomic DNA from all samples was successfully obtained. To evaluate each barcode's utility for species authentication, PCR amplification efficiency, genetic divergence, and species authentication were assessed. Among all tested regions only ITS2 locus showed 100% of PCR amplification and identification efficiencies. Based on the established method, we successfully identified two samples of Gentianae Macrophyllae Radix bought in pharmacy to the original species.

OBJECTIVETo study the effect and possible impact mechanism of salidroside on cognitive ability of Alzheimer's disease (AD) model rats induced by amyloid beta peptide (Abeta1-40).

METHODAbeta1-40 was injected into bilateral hippocampus to create the AD model. Afterwards, different doses of salidroside (25, 50, 75 mg x kg(-1)) were orally administered for 21 days. Rats' learning and memory abilities were detected by Morris water maze testing system. The levels of the superoxide dismutase (SOD), malondialdehyde (MDA), and the expression of nuclear factor-kappaB (NF-kappaB), inducible nitric oxide synthase (iNOS) and receptor for advanced glycation end products (RAGE) protein in hippocampus were also detected by different methods.

RESULTThe place navigation test showed longer escape latency, low frequency of platform quadrant crossing per unit time, damage in learning capacity, significant decrease in SOD acivity in hippocampus, notable increase in MDA content, NF-kappaB, iNOS and RAGE protein expressions in rats. Salidroside (50, 75 mg x kg(-1)) significantly alleviated the impairments of learning and memory ability. The activity of SOD increased in salidroside (50 droside group compared with that of the Alzheimer's disease group (P < 0.01).

CONCLUSIONSalidroside may treat Alzheimer's disease by inhibiting the oxidative stress.

Alzheimer Disease ; drug therapy ; Amyloid beta-Peptides ; toxicity ; Animals ; Cognition ; drug effects ; Disease Models, Animal ; Glucosides ; pharmacology ; therapeutic use ; Male ; Maze Learning ; drug effects ; NF-kappa B ; metabolism ; Nitric Oxide ; physiology ; Phenols ; pharmacology ; therapeutic use ; Rats ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; analysis ; Superoxide Dismutase ; metabolism

We used metabolomics to investigate the ability of a traditional Mongolian medicine called modified Tabusen-2 (MT-2) to improve kidney yang deficiency (KYD) in rats. All animal experiments were conducted under the guidance and standards of the Medical Ethics Committee of Inner Mongolia Medical University. SD rats were divided into 6 groups of six rats: a normal group, a model group, Jinkuishenqi pill administration group (1.26 g·kg^-1), and MT-2 administration in high-, medium- and low-dose groups (1.512, 0.756, and 0.378 g·kg^-1). KYD was established by intramuscular injection of hydrocortisone (HC) and biochemical indicators and clinical characterization was used to confirm that KYD was established. All groups received intragastrically administered drug (Jinkuishenqi pill or MT-2) or saline. Serum from each group was collected after 8 weeks and analyzed by UPLC-Q-exactive-MS to measure various biochemical indicators. The biomarkers affected by MT-2 were identified and the metabolic pathways of KYD regulated by MT-2 were analyzed by metabolomic analysis. The results show that MT-2 can decrease serum creatinine (Cr) in KYD rats and significantly increase (P<0.05) the content of thyroid stimulating hormone (TSH) and luteinizing hormone (LH). In serum samples, 38 biomarkers such as corticosterone, L-phenylalanine, and DL-tryptophan were measured as possible indicators for disease development in KYD rats. MT-2 lowered 18 biomarkers of KYD, including corticosterone, deoxycorticosterone, and L-phenylalanine, and altered 13 related metabolic pathways including phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism and steroid hormone biosynthesis, resulting in an overall improvement in KYD. MT-2 appears to be important in improving KYD in rats mainly by regulating metabolites such as amino acids, steroids and lipids.

Objective To analyze the risk factors of death in patients with subarachnoid hemorrhage induced by ruptured cerebral aneurysms and provide some therapeutic strategies. Methods A retrospective analysis of the clinical data and causes of death in 24 patients died from subarachnoid hemorrhage induced by ruptured cerebral aneurysms from January 2003 to December 2010 was performed. Results Eleven patients died from intra-cerebral re-bleeding: 5 patients had re-bleeding before the interventional embolization,3 had bleeding during the surgery,and 3 had re-bleeding after the surgery. Eight patients died from vasospasm or cerebral infarction after the surgery, including 3 with cerebral infarction under early-stage CT (within 3 d of surgery) and 5 with vasospasm or cerebral infarction under late-stage CT (3 d after the surgery); 1 died from respiratory arrest after the operation; 3 died from pulmonary infection; and 1 died from renal failure. Conclusion Re-bleeding,vasospasm/cerebral infarction and Non- neurological complications are the major causes of death in patients died from aneurysmal subarachnoid hemorrhage induced by ruptured cerebral aneurysms.Increase the awareness of causes of death will help reduce the mortality.