Three butylphthalide derivatives were isolated from the Rhizome of Ligusticum chuanxiong using a series of isolation and purification approaches including macroporous resin, ODS-A column, Sephadex LH-20 and preparative HPLC. These structures were elucidated based on extensive spectroscopic data (UV, IR, HR-ESI-MS and NMR) and identified as (3Z,3aE)-(6R,7R,2'S)-6-hydroxy-7-(2-carboxyl-2-hydroxyethylthio)-3-(2-hydroxybutylidene)-4,5,6,7-tetrahydro-phthalide (1), (3Z,3aZ)-3-butylidene-6,7-dihydroxy-4,5,6,7-tetrahydro-phthalide 7-O-α-D-glucopyranosyl-(1→2)-β-D-fructo-furanoside (2) and 3-(3-β-D-glucopyranosyloxy-butylidene)-7-hydroxy-phthalide (3).

The effect of CO2 and the manner of CO2 offer on the growth rate and maximual cell density of ultro-high density culture of Chaetoceros mulleri in the photobioreactor were studied in the work. The amount of CO2 offered to the culture was controlled by the parameter of pH value in the culture. Furthermore the growth kinetics of Chaetoceros muller in the photobioreactor was studied. The results showed requirement of CO2 by the cells and the increase of pH in the culture were the key limiting factors to the growth, when a high cell concentration in the culture was reached. The offer of CO2 could improve the statute of CO2, could control the pH in the culture and increase the growth rate and maximum cell density. The results from the experiments of CO2 offer manner showed different efficiency to growth was resulted from differences of CO2 offer manner. The best way is mixing the CO2 and air before the CO2 was offered to the culture.
Bioreactors ; Carbon Dioxide ; pharmacology ; Culture Media ; Culture Techniques ; methods ; Diatoms ; growth & development

In this study, the psbA-trnH sequence as DNA barcode was used to evaluate the accuracy and stability for identification pteridophyte medicinal material Pyrrosiae Foliumas from adulterants. Genomic DNA from 106 samples were extracted successfully. The Kimura 2-Parameter (K2P) distances and ML tree were calculated using software MEGA 6.0. The intra-specific genetic distances of 3 original plants were lower than inter-specific genetic distances of adulterants. The ML tree indicated that Pyrrosiae Folium can be distinguished from its adulterants obviously. Therefore, the psbA-trnH sequence as a barcode of the pteridophyte, can accurately and stably distinguish Pyrrosiae Folium from its adulterants.
Base Sequence ; DNA Barcoding, Taxonomic ; methods ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Ferns ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Quality Control

OBJECTIVETo know the genotype and subtype of hantavirus (HV) which infected persons in Hebei province.

METHODSAccording to G2 coding region of 76-118 and R22 strains, specific type primers were designed to detect and identity the types of HV in HFRS patients' sera with RT-nested PCR. Nucleotides were assayed from partial products after purification and reclaim. Then, gene analysis was done with DNAStar package.

RESULTS17 out of 69 positive serum specimens were successfully detected by RT-PCR and the detection rate was 24.64%, among which, or= 14 days were 0. 17 positive specimens were all belonged to SEO. The nucleotide homology of 9 typical specimens was 92.0%-100%. Between HeB7 and other 8 specimens was 92%-95%, and they belonged to different subtypes. When HeB7 compared with R22 strain, it was 97.7%. HeB7 and R22 belonged to S1 subtype. The 8 specimens except HeB7 was 95.7%-100% and they all belonged to S3 subtype. When compared with 76-118 strain, 9 specimens' nucleotide homology was only 70.3%-72.7%, belonged to different type.

CONCLUSIONSEO was the major type of HV from HFRS patients in Hebei province, S3 was the major subtype and S1 was also existed. In a certain area, the HV which belonged to the same type was correspondingly conservative, and had the characteristic of regional stability.

China ; Genotype ; Hantavirus ; classification ; genetics ; Hemorrhagic Fever with Renal Syndrome ; diagnosis ; prevention & control ; therapy ; virology ; Humans ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Viral Envelope Proteins ; genetics

OBJECTIVETo evaluate the middle term effectiveness of medial patellofemoral ligament (MPFL) reconstruction for the treatment of recurrent patellar dislocation.

METHODSFrom February 2007 to January 2010, 65 patients including 6 males and 59 females with recurrent patellar dislocation received the MPFL reconstruction. The reconstruction was performed using ipsilateral semitendinosis tendon to restore the damaged MPFL. Patients were evaluated pre-operatively and post-operatively by physical and subjectively with the IKDC (International Knee Documentation Committee), Tegner, and Lysholm questionnaires and radiographic examination.

RESULTSThe average follow-up duration was 20 months (ranged, 15 to 23 months). No recurrent episodes of dislocation or subluxation occurred. A firm endpoint to lateral patellar translation was noted in all patients at most recent follow-up. The Lysholm subjective knee evaluation score improved from (60.6 +/- 3.7) preoperatively to (89.8 +/- 4.6) postoperatively; and Tegner scores improved from (3.6 +/- 0.4) to (5.6 +/- 0.3), IKDC from (40.0 +/- 3.5) to (82.0 +/- 3.6). Radiographic evaluation demonstrated improvements in the congruence and sulcus femoral angles.

CONCLUSIONMPFL reconstruction is an effective surgical procedure for the treatment of recurrent patellar dislocation.

Adolescent ; Female ; Femur ; surgery ; Humans ; Internal Fixators ; Ligaments ; surgery ; Male ; Patella ; surgery ; Patellar Dislocation ; surgery ; Reconstructive Surgical Procedures ; Young Adult

In order to evaluate the efficiency of ITS2 and psbA-trnH sequences used as DNA barcodes to distinguish Plantaginis Semen from its adulterants, we collected 71 samples of Plantaginis Semen and its adulterants. The ITS2 and psbA-trnH sequences were aligned through Clustal W, and the genetic distances were calculated by kimura 2-parameter (K2P) model and the Neighbor-Joining (NJ) phylogenetic trees were constructed using MEGA 5.1. The results indicated that the ITS2 sequence lengths of Plantago asiatica and P. depressa were 199 bp and 200 bp, respectively; the maximum intra-specific K2P distance were lower than the minimum inter-specific K2P distance; the NJ tree based on ITS2 sequence indicated that Plantaginis Semen and its adulterants could be distinguished clearly. The sequence lengths of psbA-trnH of both P. asiatica and P. depressa were 340 bp; the maximum intra-specific K2P distances were lower than the minimum inter-specific K2P distance; the NJ tree based on psbA-trnH sequence showed that Plantaginis Semen can be distinguished clearly from its adulterants except for P. major. Therefore, ITS2 sequences can be used as an ideal DNA barcode to distinguish Plantaginis Semen from its adulterants.
Base Sequence ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Plantago ; classification ; genetics ; Quality Control ; Seeds ; classification ; genetics

OBJECTIVE Based on the methods of microdialysis,HPLC-MS/MS and gene chip tech-nology,the mechanism of Baicalin and Geniposide(BC/GP)against excitatory amino acid toxicity in ce-rebral ischemia was studied. This will provide guidance for the clinical application of BC/GP and the study of excitatory amino acid toxicity in cerebral ischemia.METHODS (1)Microdialysis technique and HPLC-MS/MS was performed to study the pharmacodynamics of BC/GP against cerebral ischemia. ①18 SD rats with body weight of(280±20)g were randomly divided into control group,treatment groups with BC/CP at low dose,medium dose and high dose(equal to the dosage of crude drugs for 30 mg·kg-1, 45 mg·kg-1and 60 mg·kg-1respectively).Rats in each group were given intragastric administration for seven days to establish cerebral ischemia model. Then, microdialysis probe was applied to collect cerebrospinal fluid from hippocampus before and after cerebral ischemia. ② First, we established the HPLC-MS/MS method for measuring drugs and excitatory amino acids.Then we detected the microdi-alysis samples and observed their changes in animals.(2)The mechanism of BC/GP against excitatory toxicity of cerebral ischemia were observed at gene level by chip technique. ① 16 SD rats with body weight of 240±20 g were randomly divided into sham group, model group, treatment group of BC(60 mg·kg-1),treatment group of GP(60 mg·kg-1)and treatment group of BC/GP(7:3)(60 mg·kg-1).Rats in eachgroup were given intragastric administration for seven days to establish cerebral ischemia model. Then the rats were sacrificed,and the hippocampus were rapidly harvested and stored at-80℃for further detection. ②After the quality inspection of the hippocampal,the qualified samples were subjected to detect the levels of neurotransmitter receptor gene in the ischemic of rats by gene chip technology.Finally,the results were analyzed by the method of Δ ΔCt.RESULTS (1)Only three compounds includ-ed GP,glutamic acid and aspartic acid were detected in microdialysis samples by HPLC-MS/MS.The concentration of GP increased and lasted for 120 min with a significant dose-dependent after cerebral ischemia.Compared with low dose group,the AUC(0-t),MRT(0-∞),Cmaxand t1/2zin high-dose group showed significant difference(P<0.01).Compared with the model group,the levels of glutamic acid and aspartic acid in the treatment groups decreased significantly,especially in the middle and high dose groups.(2) 89 genes in the neurotransmitter receptor gene signaling pathway were detected by gene chip technol-ogy. There were 22 genes with |Fold Regulation|>1.5 in the model group, compared with the sham group.Five of the 22 genes showed statistically significant differences,including Grin2c(2.9026),Chrna7 (-1.5877), and Tacr2 (-1.7695). Htr3a (-1.8172) and Grm6 (-2.3527). There were 5 genes with |Fold Regulation|>1.5 in the BC group, compared with the model group, Two of them exhibited statistically significant differences,including Brs3(1.797)and Grin2c(-1.7979).There were 14 genes with|Fold Reg-ulation|>1.5 in the GP group, compared with the model group. Three of them displayed statistically significant differences,including Hcrtr2 (-1.6584), Sctr (-3.8524) and Grin2c (-4.8408). Compared with model group, the genes of |Fold Regulation|>1.5 in BC/GP (7:3) group are 5, and only one of them showed a significant differences. CONCLUSION (1)After administration of BC and GP,GP can cross the blood-brain barrier and reduce the release of excitatory amino acids in the hippocampus. (2) BC/GP can inhibit the interaction between excitatory amino acids and excitatory amino acid receptors and attenuate the toxicity of excitatory amino acids by down-regulating the expression of glutamic acid receptor Grin2c gene.(3)BC/GP may exert their brain protection effect by reducing the release of excit-atory amino acids and inhibiting the expression of excitatory amino acid receptors.

OBJECTIVETo analysis the effect of amiodarone on funny current (I(f)) and hyperpolarization-activated cation channel (HCN) gene expressions of the neonatal rat ventricular myocytes.

METHODSVentricular myocytes of 1 - 3 days-old rats were isolated and cultured. The cardiomyocytes were treated by amiodarone (0.01, 0.1, 1, 10, 100 micromol/L) for 3 hours or amiodaron (10 micromol/L) for 0, 0.5, 1, 3, 6 hours. The I(f) and HCN 1 - 4 gene expressions were measured through the whole-cell configuration of the patch-clamp technique and real-time quantitative polymerase chain reaction (real-time PCR) using SYBR Green PCR kit.

RESULTS(1) HCN1, HCN2, HCN3 and HCN4 represented (0.23 +/- 0.01)%, (83.58 +/- 0.04)%, (0.79 +/- 0.01)% and (15.44 +/- 0.01)% of total HCN mRNA, respectively. (2) Amiodaron resulted in a dose-dependent I(f) [(3.1 +/- 0.9)%, (9.7 +/- 2.4)%, (36.7 +/- 5.8)%, (80.3 +/- 1.8)% and (85.9 +/- 3.1)%, respectively at -145 mV, IC(50) (1.32 +/- 0.28) micromol/L], HCN2 [(2.1 +/- 0.8)%, (8.9 +/- 3.6)%, (30.1 +/- 4.2)%, (78.3 +/- 3.6)% and (81.1 +/- 1.9)%, respectively] and HCN4 decrease [(0.5 +/- 0.2)%, (2.1 +/- 2.6)%, (8.8 +/- 3.2)%, (60.1 +/- 4.6)% and (59.6 +/- 6.5)%, respectively]. (3) Amiodaron (10 micromol/L) also induced a time-dependent I(f) [(1.1 +/- 0.1)%, (12.6 +/- 2.3)%, (80.6 +/- 2.2)% and (80.1 +/- 2.1)%, respectively], HCN2 [(1.0 +/- 0.1)%, (9.8 +/- 3.9)%, (82.9 +/- 4.6)% and (83.9 +/- 1.7)%, respectively] and HCN4 decrease [(0.1 +/- 0.1)%, (1.9 +/- 1.1)%, (59.4 +/- 7.8)% and (60.9 +/- 3.1)%, respectively]. However, HCN1 and HCN3 expressions were not affected by amiodaron treatment.

CONCLUSIONCurrent density of I(f) and the expression of HCN2 and HCN4 were decreased by amiodaron which might be the possible antiarrhythmic working mechanisms of amiodaron.

Amiodarone ; pharmacology ; Animals ; Animals, Newborn ; Cells, Cultured ; Cyclic Nucleotide-Gated Cation Channels ; genetics ; metabolism ; Female ; Gene Expression ; Heart Ventricles ; metabolism ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels ; Male ; Myocytes, Cardiac ; drug effects ; metabolism ; Patch-Clamp Techniques ; Potassium Channels ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate the clinical value of sweat function examination in early diagnosis of diabetic peripheral neuropathy (DPN).

METHODSNinety-eight hospitalized type 2 diabetic patients with or without DPN (DN and DC groups) according to Michigan Diabetic Neruopathy Score (DNS) and 40 healthy volunteers (NC group) were evaluated for their sweat function of the feet in relation to the peripheral autonomic nerve with sweat printing method using Neuropad. The Neuropad color-changing time was recorded to assess the sensitivity and specificity of sweat printing methods relative to DNS for DNP evaluation, and the correlation of the Neuropad color-changing time to DNS score was analyzed.

RESULTSThe average Neuropad color-changing time was 4.0-/+0.6, 4.3-/+1.2 and 23.0-/+6.1 min in NC, DC, and DN groups, respectively, showing significant differences between the 3 groups (P<0.05). The morbidity rate detected by sweat printing method was 62.2%, similar to that detected by DNS (57.1%, P>0.05). The sensitivity of the sweat printing method for DPN diagnosis was 92.8%, with specificity of 78.5%, positive predictive value of 93.2%, and negative predictive value of 78.6%. DNS showed significant positive correlation with the Neuropad color-changing time (r=0.46, P<0.05).

CONCLUSIONSweat printing method provides an objective, simple and reliable method for sweat function evaluation of the feet of type 2 diabetic patients to help in early DPN diagnosis, and quantification of the results of sweat printing method can be indicative of the DPN severity.

Case-Control Studies ; Color ; Diabetic Neuropathies ; diagnosis ; physiopathology ; Early Diagnosis ; Foot ; physiopathology ; Humans ; Male ; Middle Aged ; Sweating ; physiology ; Time Factors

AIM To study the effects of Yizhi Congming Decoction (Ginseng Radix et Rhizoma,Poria,Polygoni multiflori Radix praeparata,etc.) on learning and memory deficits through reducing tau hyperphosphorylation in Aβ-induced AD mice and its mechanism of action.METHODS Sixty ICR mice were randomly and equally assigned into six groups.Sham group (injection of 3 μL normal saline into left hippocampus),model group,donepezil group and each group of Yizhi Congming Decoction (injection of 3 μL Aβ25-35 into left hippocampus).Donepezil group and three treatment groups were gavaged with donepezil and Yizhi Congming Decoction.After 15 days of administration,immunohistochemistry was used to observe the expressions of Ser404 and Thr231.Western blot was performed to evaluate the levels of phosphatidylinositol-3-kinase (PI3K),threonine/serine protein kinase B (AKT) and glycogen synthase kinase 3β (GSK3β).RESULTS Yizhi Congming Decoction attenuated the levels of phosphorylated tau at the Thr231 and Ser404 sites in the hippocampus and increased the phosphorylation levels of PI3K,AKT and GSK3β.CONCLUSION Yizhi Congming Decoction effectively provides protection against learning and memory deficits and inhibits hyperphosphorylated tau protein expression in the hippocampus.The possible mechanism may occur via the PI3 K/Akt-dependent GSK3 β signalling pathway.