This study aimed to identify Belamcandae Rhizoma, Iridis tectori Rhizoma and their adulterants by ITS2 sequence. All the DNA samples of Belamcandae Rhizoma, Iridis tectori Rhizoma and their adulterants were extracted. The ITS2 sequence were succesfully amplified, and purified PCR products were sequenced. All the sequences were assembled using the CondonCode Aligner V3.7.1. The Kimura 2-Parameter (K2P) genetic distances and the Neighbor-Joining (NJ) phylogenetic tree were calculated by using MEGA5.1. Results indicated that the maximum intraspecific genetic distances of Belamcandae Rhizoma was 0, and the average GC content was 52.22%;the maximum intraspecific genetic distances of Iridis tectori Rhizoma was 0.004, and the average GC content was 67.87%. The maximum K2P intraspecific genetic distance of Belamcanda chinensis, Iris tectorum were both lower than the minimum interspecific genetic distance of adulterants. Additionally, the ITS2 sequences in each of these polytypic species were separated into pairs of divergent clusters in the NJ tree. The NJ tree based on ITS2 sequence indicated that Belamcandae Rhizoma, Iridis tectori Rhizoma and their adulterants could be distinguished clearly. It could be concluded that ITS2 barcode can be used to correctly identify Belamcandae Rhizoma, Iridis tectori Rhizoma from their adulterants, and ensure their safety in use.

Objective To clarify the effects of simulated microgravity on the local regulating function of the pulmonary circulation. Method Rat tail-suspension(TS) was used as the animal model to simulate the effects of microgravity.The changes in responses of pulmonary artery to several vasoactive agents were determined by using in vitro vessel rings perfusion technique. Result As for the endothelium-intact vessel rings from TS group compared with those from the control(C) group,the contractile responses of pulmonary artery to potassium chloride (KCl,10～70 mmol/L) did not change obviously after 7 d tail-suspension,which were markedly diminished as to phenylephrine (PE,10-9～10-5mol/L).The vasodilatory responses to acetylcholine (ACh,10-9～10-5mol/L) were markedly enhanced,which did not change obviously as to captopril (Cap,10-5～3×10-3 mol/L).The vasoreactivity of endothelium-removed pulmonary artery rings to all vasoactive agents did not change after 7 d tail-suspension as compared with those from control rats. Conclusion The release of nitric oxide (NO) by rat pulmonary arterial endothelium was enhanced after 7 d tail-suspension.The vasodilatory function of pulmonary artery was enhanced.

Objective To evaluate the effect of surgical treatment for primary hepatocellular carcinoma(HCC) with bile duct thrombi(BDT).Methods Eleven cases were diagnosed as HCC with BDT by B ultrasound,CT,MRCP or ERCP.Hepatectomy and choledochotomy were performed,resecting the primary tumor and cleaning the BDT completely,followed with TACE.Results No patients died during operation.The survival time of 9 patients who received hepatectomy and BDT clearance ranged from 3 to 38(average 18.5) months,and that of the other two patients received TACE and BDT clearance survived 6 and 11 months respectively.Conclusion Accurate diagnosis and radical hepatectomy with complete BDT clearance are effective treatment for HCC with BDT.

Objective To study the expression of Rho kinase and its functional activation in lung tissue from hypoxic pulmonary hypertension(HPH) rat model,and the effects of fasudil on HPH.Methods Seventy-two male Spraque-Dawley rats were randomly divided into the control group,hypoxic model group,and fasudil-intervention group[group with hypoxia and fasudil for 15 mg/(kg?d)],respectively.Mean pulmonary arterial pressure(mPAP) and right ventricle hypertrophy index(RVHI) were measured.Expression of Rho kinase mRNA and protein were examined by reverse transcriptase-polymerase chain reaction(RT-PCR) and Western blot,respectively.The phosphorylation of binding subunit of myosin phosphatase(MBS)-a substrate of Rho kinase was detected by Western blot and defined,as the mark of functional activation of the kinase.Results The expression of Rho kinase mRNA in hypoxic model group was markedly upregulated even before the onset of the third day after the experiment(HPH),and it was much lower in rats of fasudil group than that of hypoxic model group.The phosphorylation of MBS was significantly higher in hypoxic model group than that in control group,and it was positively correlated with the mPAP and RVHI(all P

Objective To observe the effect of acupuncture combined with integrated Chinese-Western neuromuscular facilitation technique on persistent vegetative state (PVS).Methods41 PVS cases were randomly divided into the observation group (n=21) and control group (n=20). The observation group was treated with acupuncture, neuromuscular facilitation technique combined with reasonable obligatory exercise, neuromuscular electrical stimulation, manipulation treatment and Chinese medicine. The control group was treated with the Western and Chinese medicines, and hyperbaric oxygen.ResultsAfter 1～3 months treatment, 7 cases cured,8 cases were markedly effective, 5 cases were effective, 1 cases were ineffective in the observation group with a total markedly effective rate (71.4%) and effective rate (95.2%). While, in the control group, 3 cases cured, 5 cases were markedly effective, 6 cases were effective, 6 cases were ineffective and total markedly effective rate was 40%, effective rate was 70%. There was a significant difference between the two groups in the total markedly effective rate and the effective rate ( P<0.05). The average PVS score increased by 7.46±1.22 in the observation group and 4.59±1.21 in the control group. Also there was a significant difference between the two groups ( P<0.001).ConclusionThe therapy of acupuncture combined with integrated Chinese-Western neuromuscular facilitation technique can markedly promote PVS patients coming round and improve patients' prognosis.

To explore a new method to identify Moutan Cortex to guarantee its safe use, internal transcribed spacer 2 (ITS2) sequence was used to identify Moutan Cortex and its adulterants. DNA was extracted and target fragments were amplified. Sequences were analyzed and assembled by CodonCode Aligner V3.7.1. Genetic distances were computed and phylogenetic tree was constructed based on kimura 2-parameter (K2P) model by MEGA 5.0. The length of the 20 ITS2 sequences of Moutan Cortex from nine different places is 227 bp, and no variation site was detected. The maximum inter-specificK2P distance of Moutan Cortex is 0, the minimum intra-specific K2P distance is 0.041, the average intra-specific K2P distance is 0.222. According to NJ analysis, Moutan Cortex from different places can get together as one branch with bootstrap support values 99%, which indicates Moutan Cortex can be easily distinguished from its adulterants. Using ITS2 sequence can accurately identify Moutan Cortex and its adulterants, it is an effective supplementary to traditional identification methods.
Base Sequence ; China ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Paeonia ; classification ; genetics ; Phylogeny ; Quality Control

The objective of the present study was to establish a method based on principal component analysis (PCA) for the study of transdermal delivery of Chinese medicinal formulae, and to choose the best penetration enhancers for Qingfei Xiaocuo gel depend on this method. Using improved Franz type diffusion cell and excised rat skin in vitro as transdermal barrier, the receptive solution fingerprint was established by HPLC, harvesting the areas of the common peaks in the fingerprint, then the total factor scores of the concentrations at different times were calculated using PCA and were employed instead of the concentrations to compute the cumulative amounts (Q12) and enhancement ratio (ER), the latter of which were considered as the indexes for optimizing penetration enhancers. Compare to the control group, the ER of the other groups increased significantly and furthermore, 2.5% azone with 2.5% menthol manifested the best effect. PCA represent most information in the receptive solution, the method above could choose the best penetration enhancers, it could be a reference for the study of transdermal delivery of Chinese medicinal formulae.
Administration, Cutaneous ; Animals ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Gels ; In Vitro Techniques ; Male ; Medicine, Chinese Traditional ; Mice ; Principal Component Analysis ; Skin ; metabolism

OBJECTIVETo study the effect of nano-SiO2 on spatial learning and memory.

METHODSTwenty-four male rats were randomly divided into 3 groups: control group (C group), low dose group (L group) and high dose group (H group). The rats were intragastrically administrated with nanometer particles at 25 and 100 mg/kg body weight every day for 4 weeks. After exposure, the ability of learning and memory of rats was tested by Morris water maze, and electrophysiological brain stereotactic method was used to test long-tear potentiation (LTP) in dentate gyrus (DG) of the rats.

RESULTSThe increase rate of body weight in H group was reduced significantly compared with C group ( P < 0.05). In the space exploration experiment of Morris water maze test, the escape latency of H group was longer than that of C group (P < 0.05). The rats of H group spent less time in finding the target quadrant (P < 0.05) . The rate of LP induction of H group was significantly lower than that of C group (P < 0.05). After high fre quency stimulation (HFS), The changes of amplitude of population spike (PS) of L group and H group were lower than those of C group significantly (P < 0.05, P < 0.01).

CONCLUSIONNano-SiO₂may result in impairment of spatial learning and memory ability by reducing the rate of LTP induction and the increase of PS in hippocampus.

Animals ; Dentate Gyrus ; drug effects ; Long-Term Potentiation ; drug effects ; Male ; Maze Learning ; drug effects ; Memory ; drug effects ; Nanoparticles ; adverse effects ; Rats ; Silicon Dioxide ; adverse effects ; Spatial Learning ; drug effects

Objective To Investigate the influence of high-intensity pulsed electromagnetic field (HIPEMF) on neural differentiation of neonatal rats neural stem cells in vitro.Methods Neural stem cells (NSCs) were isolated from the subventricular zone of 3-day-old neonatal rats and cultured with serum-free condition medium for 14 days.All the NSCs were then randomly classified into an experimental group which received the stimulation of HIPEMF (0.1 Hz,4 T,8 pulses) and a control group which received no special intervention.Differentiation of the culture was induced by addition of 10％ fetal bovine serum on the first day after intervention,the morphological changes of cells were observed under the microscope at different time points.The NSCs adhered to the wall and differentiated for seven days,the immunofluorescence was employed to observed and calculate the ratio of differentiated cells with astrocyte marker GFAP or neuronal markers TUJ1.RT-PCR and western blotting were used to measure the expression levels of the differentiated cells based on gene and protein levels,respectively.Results Immunofluorescence staining showed the number of TUJI positive cells in the experimental group(33.4％ ± 5.1％)was significantly more than the control group (26.5％ ± 7.0％),while the number of GFAP positive cells was decreased(23.9％ ± 5.0％) as compared with the control group(36.2％ ± 2.2％).RT-PCR showed that the TUJ1 mRNA expression levels in the experimental group was (1.682 ± 0.086) times of the control group.Western blot showed that the expression of TUJ1 (0.729 ±0.061) in the experimental group was higher than in the control group (0.590 ± 0.157),while the expression of GFAP in the experimental group (0.566 ± 0.056) was less than in the control group(1.034 ± 0.051).Conclusions HIPEMF facilitates differentiation of neural stem cells to neurons,at the cost of reducing astrocytic differentiation.

Objective To explore the related risk factors of cerebral hemorrhage complicated with stress ulcer (SU). Methods The clinical data of 1 185 patients with cerebral hemorrhage admitted to Department of Emergency Medicine of Nanjing General Hospital from March 2006 to March 2014 were retrospectively analyzed. Patients were divided into two groups according to whether patients complicated with SU or not. Data was collected within 8 hours after admission in two groups including gender,age,amount of bleeding,the bleeding site (basal ganglia,thalamus, brainstem,brain lobe,ventricle,subarachnoid,and cerebellum),disturbance of consciousness,acute physiology and chronic health evaluationⅡ(APACHEⅡ)score,systolic blood pressure(SBP),history of hypertension,and history of cerebral hemorrhage. The statistically significant risk factors found using univariate analysis was selected and was analyzed to find independent risk factors with multivariate logistic regression analysis. The receiver operating characteristic curve (ROC curve)was plotted to analyze the independent risk factors and evaluate their power of test. Results 1 185 patients with cerebral hemorrhage were enrolled in the study,293 cases occurred SU,accounting for 24.7%,and 892 cases without SU,which accounted for 75.3%. As shown by univariate analysis,risk factors for cerebral hemorrhage complicated with SU included age,amount of bleeding,the bleeding site,disturbance of consciousness,APACHEⅡscore,SBP. As to the site of bleeding,brain,thalamus,brainstem hemorrhage complicated with SU were higher proportion,45.3%(43/95),39.1%(63/161),36.9%(48/130),which were significantly higher than those of the lobes of the brain 〔26.2% (33/126)〕,cerebellum 〔18.8% (15/80)〕,basal ganglia〔16.1%(78/485)〕,arachnoid the inferior vena cava 〔12.0% (13/108)〕. Multivariate logistic regression analysis showed that amount of bleeding 〔odds ratio (OR)=3.305,P=0.001,95%confidence interval (95%CI)2.213-48.634〕,the bleeding site (OR=1.762,P=0.008,95%CI 0.123-2.743),SBP (OR=1.223,P=0.034,95%CI 0.245-2.812) were independent risk factors of cerebral hemorrhage complicated with SU. The area under the ROC curve (AUC)of amount of bleeding and SBP were 0.846 and 0.597,suggesting that amount of bleeding has moderate diagnostic value and SBP has low diagnostic value. Conclusions Cerebral hemorrhage patients with large amount of bleeding,the bleeding site in the ventricle,thalamus or brainstem,high SBP are of great risk. We should lower blood pressure and give preventive treatment for SU as soon as possible.