Objective To obtain the indispensable key enzyme-hydroxide methyl enylamino 4-cyclodiphosphate synthase (HDS) gene involved in the MEP pathway cloned from Artemisia annua and conduct bioinformatic and functional complementation analysis. Methods To perform multiple sequence alignment for the nucleotide acid sequence of the other reported seed plants' HDS gene, to select conservative areas for designing degenerate primers, and to gain the aim gene from A. annua through homologous expanding and cDNA bottom speedily expanding technique. To perform sequence alignment using BLAST, to identify open reading frame (ORF) using ORF Finder, and to construct phylogenetic tree using neighbor joining (NJ) ways in MEGA3.0. Results The obtained HDS cDNA sequence was 2 324 bp containing a 1 854 bp ORF and encoding a 617-amino acid protein. Bioinformatic analysis showed that AaHDS was homologous with HDS derived from other seed plant species. Functional complementation analysis indicated that AaHDS could make up the short HDS function of mutant Escherichia coli MG1655 ara<>HDS. It could make the mutant get back to upgrowth, which showed AaHDS had typical HDS gene function. Conclusion The cloning HDS gene from A. annua for the first time provides a good basis for further study on the metabolization project of artemisinin.

ObjectiveTo observe the effects of hypoxia inducible factor-1 alpha (HIF-1α)-targeting small interfering RNA(siRNA) on the expression of HIF-1α and vascular endothelial growth factor (VEGF) in HaCaT ceils under hypoxic conditions. MethodsHaCaT cells were cultured and divided into four groups, normal control group (without any treatment), hypoxia group (cultured under hypoxic conditions for 24 hours),liposome control group (transfected with liposome followed by hypoxic culture for 24 hours), RNA interference group (transfected with HIF-1α-targeting siRNA/liposome complexes followed by hypoxic culture for 24 hours). Fluorescence real-time quantitative PCR was utilized to determine HIF-1oα and VEGF mRNA expression in HaCaT cells, and Western blot to detect HIF-1α and VEGF protein expression. ResultsNo significant difference was observed in the mRNA expression of HIF-1α between the hypoxia group and normal control group(0.907 ± 0.032 vs. 0.878 ± 0.034, F =1.108, P ＞ 0.05), while the expression levels of VEGF mRNA,HIF-1α and VEGF proteins were significantly higher in the hypoxia group than in the normal control group (0.935 ± 0.032 vs. 0.652 ± 0.053, 0.813 ± 0.047 vs. 0.236 ± 0.014, 0.791 ± 0.030 vs. 0.316 ± 0.013, all P ＜0.05). A significant decline was noted in the mRNA and protein expression levels of VEGF (0.230 ± 0.044 vs.0.978 ± 0.030, 0.213 ± 0.026 vs. 0.817 ± 0.049, both P ＜ 0.05) and HIF-1α(0.497 ± 0.033 vs. 0.806 ±0.040, 0.249 ± 0.028 vs. 0.833 ± 0.052, both P ＜ 0.05) in the RNA interference group than in the liposome control group. ConclusionsHypoxia may enhance the expression of HIF-1α and VEGF in HaCaT cells, and to inhibit the HIF-1α expression may suppress the expression of VEGF in HaCaT cells under hypoxia.

Objective To investigated the serum level of carcinoma antigen 125 (CA-125) and its clinical significance in patients with Budd-Chiari syndrome.Methods We reviewed medical records and laboratory tests of patients with BCS first diagnosed in our hospital between August 2011 and April 2013.235 patients were included as experiment group,while 120 healthy adult volunteers were randomly selected as control group.The serum level of CA-125 were detected by electrochemilumescence immunization assay in this single-center retrospective control study.Results The average serum level of CA-125 in experiment group is higher than that of control group [(147.9 ±246.6) kU/L vs (16.0 ±7.2) kU/L,P ＜0.001].In experiment group,the relative coefficient for serum CA-125 with ascites,alanine aminotransferase,aspartate aminotransferase,albumin and Rotterdam BCS scores was 0.79,0.45,0.29,-0.393 and 0.71,respec tively,P ＜0.001.As of October 2013,we found that the 68 BCS patients with serum CA-125 level 5-fold higher than the upper limit of normal (＞ 175 kU/L) presented much lower survival rates and asymptomatic survival rates than the rest 167 BCS patients after intervention therapy:(95.6％ and 79.8％) vs (98.8％ and 92.0％),P ＜ 0.05.Conclusions The serum level of CA-125 in BCS patients have positive correlation with ascites volume,liver injury degree and Rotterdam BCS scores.Serum CA-125 evaluation appears to be a valuable examination option in BCS as CA-125 levels negatively correlate with worse prognosis,thus could be applied as an efficient tool for prognostication.

Objective To study the clinical features and prognosis of patients with primary BuddChiari syndrome (BCS) caused by hepatic vein thrombosis.Method 16 patients with primary BCS caused by hepatic vein thrombosis treated in our hospital between June 2010 to December 2012 were used as the study group while 132 patients with primary BCS caused by other causes were used as the control group.A retrospective study was then employed to analyze the clinical data of the two groups of patients during hospitalization and on follow-up.The study was censored in June 2013.The median follow-up was 24 months (range,6 months to 36 months).The difference in quantitative data between the 2 groups were analyzed using the independent-samples t test and the Wilcoxon W rank sum test,and the difference in qualitative data were analyzed using the Chi-square test and the Fisher's exact test.The survival rates and recurrence rates were calculated using the Kaplan-Meier method.Result The study group was significantly lower than the control group in age,duration of symptoms,albumin level,diameter of spleen and survival rate,but it was significantly higher in the proportion of patients with ascites,average hospitalization time,alanine transaminase,aspartate aminotransferase,total bilirubin,carbohydrate antigen-125 and recurrence rate after percutaneous transluminal angioplasty.The differences were significant (P ＜ 0.05).The Rotterdam BCS prognosis grades of the study group were:9 patients grade Ⅱ and 7 patients grade Ⅲ.In the control group,there were 65 patients with grade Ⅰ,51 patients with grade Ⅱ,and 16 patients with grade Ⅲ.The prognosis grade of the study group was significantly higher than the control group (P ＜ 0.05).Conclusion When compared to the patients with BCS due to other causes,patients with BCS caused by hepatic vein thrombosis were more common in the young,most of them were diagnosed in the acute period,they had worse clinical outcomes and had more severe clinical symptoms and liver damage.

By Silica gel, Sephadex LH-20 and other materials for isolation and purification and by physicochemical methods and spectral analysis for structural identification, 32 compounds were isolated and identified from ethyl acetate portion of alcohol extract of the Osmanthus fragrans. Their structures were identified as boschniakinic acid (1), ursolaldehyde (2), augustic acid (3), arjunolic acid (4), 5-hydroxymethyl-2-furancarboxaldehyde (5), isoscutellarein (6), 6, 7-dihydroxycoumarin (7), 2α-hydroxy-oleanolic acid (8), quercetin-3-0-β-D-glu-copyranoside (9), D-allito (10), 5, 4'-dihydroxy-7- methoxyflavone-3-0-β-D-glucopyranoside (11), 5,7-dihydroxychromone (12), lupeol (13), naringenin (14), acetyloleanolic acid (15), chlorogenic acid (16), kaempferol-3-0-β- D-glucopyranoside (17), oleanolic acid (18), kaempferol-3-0-β-D-galactopyanoside (19), 3', 7-dihydroxy-4'-methoxyisoflavon (20), ergosta-4,6,8 (14), 22-tetraen-3-one (21), p-hydroxycinnamic acid (22), syringaresinol (23), 3,4-dihydroxyacetophenonel (24), β-sitosterol (25), ethyl p-hydroxyphenylacetate (26), benzoic acid (27), caffeic acid (28), coelonin (29), p-hydorxy-phenylacetic acid (30), p-hydroxyacetophenone (31), and methyl-p-hydroxphenylacetate (32). Except for compounds 2, 4, 5, 8-11, 13, 15, 18, 20, 25, and 27, the rest were isolated from the Osmanthus fragrans for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Oleaceae ; chemistry ; Plants, Medicinal ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Hemorrhoids ; Humans ; Male ; Mucous Membrane

Currently, the herbal prescription therapy for corresponding constitutional diseases is a common constitution regulating method. This method has an obvious effect in treating and regulating constitution-related diseases. However, for people who do not have disease, they prefer to regulate constitution with dietary therapy. In this paper, the researchers came up with a design method of constitution regulating and healthcare foods based on medicinal property combination mode of clinical empirical formulas. With "Yupinfeng San", a common formula for Qi-insufficiency constitution and specific endowment constitution, as the example for constitution regulating and healthcare foods, the researchers proved the effectiveness and rationality of healthcare food schemes in terms of the efficacy of single herb and the modern pharmacological study.
Diet ; Diet Therapy ; Humans ; Plants, Medicinal ; chemistry ; metabolism ; Prescriptions

OBJECTIVE: To observe the effect of hypoxia inducible factor -1α (HIF-1α) small interfering RNA (siRNA) on the expression of HIF-1α and inducible nitric oxide synthase (iNOS) in HaCaT cells under hypoxia. METHODS: HaCaT cells were divided into 4 groups: the normal control group (without any treatment), the hypoxia group (under hypoxia for 24 h), the liposome control group (HaCaT cells transfected with liposome before hypoxia treatment), the RNA interference group (HaCaT cells transfected with siRNA sequences then under hypoxia for 24 h). Real-time PCR and Western blot were utilized to determine HIF-1α and iNOS mRNA and protein expression in HaCaT cells. RESULTS: There was no significant difference of the mRNA expression of HIF-1α between the hypoxia group and the normoxia group (P>0.05), but the protein expressions of HIF-1α was increased in the hypoxic group than that in the normoxia group (P<0.05). Both the mRNA and protein expression of iNOS were increased in hypoxic conditions than that in the normoxia (P<0.05). Decreases were more significant in the mRNA and protein expression of HIF-1α and iNOS in the RNA interference group than that in the liposome control group in HaCaT cells (P<0.05). CONCLUSION Hypoxia increased HIF-1α and iNOS expression in HaCaT cells and inhibition of HIF-1α expression decreased iNOS expression.
Cell Hypoxia ; Cell Line ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Keratinocytes ; cytology ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics

Danggui, Agelicae Sinensis Radix, is a widely used Chinese herb to enrich blood, but its quality cannot be effectively assessed by the known chemical markers such as ferulic acid, ligustilide, polysaccharides, etc. A new bioassay was therefore developed to quantify the Enrich-Blood Bioactivity (EBB) for the quality assessment of Danggui raw materials. Danggui sample was first extracted with ethanol and water, respectively. Then the ethanolic extract and water extract were mixed as a test sample to quantify the amount of EBB by mice experiment. The blood deficiency mode in mice was developed by intraperitoneal injecting cyclophospharmide and phenylhdrazine hydrochloride. The quantity of red blood cell was chosen as EBB marker. Cyclosporine A was chosen as a control substance. EBB in analytes was quantified by the amount reaction of parallel line analysis (3, 3') method. The results indicated that the reliability test for quantifying EBB was passed through and the measured value was valid. The analytes showed the significant EBB (P < 0.05). The correlation coefficient was 0.9984 (n=5) between the amount of cyclosporine A (0.035-0.56 g x kg(-1)) and the increased number of red blood cell. The relative standard deviation (RSY) on the amount of EBB was estimated to be 6.15% (n = 6) by six replicated tests, and the confidence limit rate was 26.68% (n = 6). Five Danggui samples, which were collected from different cultivation areas with various morphological characters, showed the variety of EBB in the range of 21.95-44.16 U x g(-1). It is concluded that the developed method is accurate to quantify the EBB of Danggui raw materials, and is therefore suitable to assess its quality.
Angelica sinensis ; chemistry ; Animals ; Biological Assay ; methods ; Drugs, Chinese Herbal ; pharmacology ; Erythrocyte Count ; Erythrocytes ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Plant Roots ; chemistry

Objective To evaluate the protective effect of Yersinia pestis capsular antigen F1 and recombinant rV270 on mice after immunization with them.Methods According to body weight,40 female Balb/c mice aged 6 to 8 weeks were randomly divided into four experimental groups(Fl-10 μg + aluminum adjuvant,F1-20 μg + aluminum adjuvant,rV-10 μg + aluminum adjuvant,and rV-20 μg + aluminum adjuvant) and a control group,8 in each group.Mice in experimental groups were immunized with the natural antigen F1 and recombinant antigen rV270 adsorbed to 25％ aluminum adjuvant and the control group was immunized with the same amount of aluminum adjuvant.Each mouse was immunized at the hind leg muscle with 100 ml immunizing agent,then a booster immunization was done once on the 21st day after the first immunization.The blood of all mice was collected on the 8th week after the first immunization,serum antibody titers were detected by ELISA and the data of antibody titers were analyzed by t test for comparison between groups.At the same time the mice were injected subcutaneously with 2000-fold LD50 of Yersinia pestis virulent strain 141,after 14 days,the protective effect of immunization was analyzed.Results The control group did not produce antibody.Antibody geometric mean titers (GMT) of the F1-10 mg + aluminum adjuvant and F1-20 mg + aluminum adjuvant groups were 1 ∶ 30443.9,and 1 ∶21527.8,respectively,and compared between the two groups,the difference was not statistically significant (t =1.1282,P ＞ 0.05).The GMTs of the rV-10 μg + aluminum adjuvant and rV-20 μg + aluminum adjuvant groups were 1 ∶ 13957.3 and 1 ∶18100.9,respectively,and compared between the two groups,the difference was not statistically significant(t =0.9408,P ＞ 0.05 ).After subcutaneous injection with Yersinia pestis virulent strain 141,all mice died in the control group but all survived in the experimental group.Conclusion The immune activity of natural antigen F1 and recombinant rV270 is high,which can be used as the main component of subunit vaccine in the plague subunit vaccine study.