An investigation on the chemical constituents of the 90% EtOH extract of Perovskia atriplicifolia led to the isolation of fifteen compounds from the EtOAc fraction. Based on the detailed spectral analysis (MS, 1D and 2D NMR), as well as comparison with the literatures, the structures of compounds 1-15 were determined as cirsimaritin (1), salvigenin (2), syringaldehyde (3), vinyl caffeate (4), 2α, 3α-dihydroxyolean-12-en-28-oicacid (5), 2α, 3α-dihydroxyurs-12-en-28-oicacid (6), niga-ichigoside F1 (2α, 3β, 19α, 23- tetrahydroxyurs - 12-en-28-oicacid- O-β-D- glucopyranoside, 7), sericoside (8), 4-epi-niga-ichigoside F1 (2α, 3β, 19α, 24-tetrahydroxyurs-12-en-28-oicacid O-β-D-glucopyranoside, 9), 2α, 3β, 24-trihydroxyolean-12-en-28-oicacid O-β-D-glucopyranosyl-(1 --> 2) - β-D-glucopyranoside (10), pruvuloside A (11), asteryunnanoside A [2α, 3β, 23-trihydroxyolean-12-en-28-oicacid O-β-D-glucopyranosyl-(1 --> 2)-β- D- glucopyranoside,12], rosmarinic acid methyl ester (13), β-sitosterol (14), and daucosterol (15), respectively. Compounds 1-13 were isolated from the Perovskia genus for the first time. All the compounds were obtained from P. atriplicifolia for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Lamiaceae ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

Objective To compare the diagnostic values of magnetic resonance elastography (MRE) and T1ρ imaging in staging hepatic fibrosis (HF) in a rabbit model.Methods The institutional animal care and use committee approved all experiments.Sixty healthy rabbits were divided into HF group (n=44) and control group (n=16).Each eight rabbits in the HF group and 4 rabbits in the control group were randomly selected at the 4th,5th,6th week and the remaining rabbits at the 10th week after subcutaneous injection with 0.1 ml 50％ CCl4 oily solution per kilogram of body respectively,to undergo liver MR scan including axial liver MRE and T1ρ imaging.The values of liver stiffness (LS) and T1ρ were measured.Masson trichrome staining of liver tissue was used.According to the Scheuer scoring system,rabbits were classified into F0 to F4 group based on the percentage of hepatic fibrosis.The difference of LS values and Tip values among stage F0 to F4 were compared by the one-way ANOVA analysis.The correlations between pathological staging and LS,T1ρ values were performed by the Spearman correlation analysis.ROC curve analysis was performed to compare the value of MRE with T1ρ imaging.Results Forty three rabbits were included,there were 10,8,8,8,9 rabbits in F0,F1,F2,F3 and F4 stage,respectively.LS values were (1.051±0.155),(1.335±0.235),(1.401±0.163),(2.001±0.499) and (2.981±0.714) kPa in F0,F1,F2,F3 and F4,respectively,while T1 p values were (23.20±4.02),(24.28±2.93),(25.40± 1.82),(24.69± 1.85) and (31.54±3.39) ms (all P＜0.05).The correlation of LS values with hepatic fibrosis staging measured on MRE was stronger than T19 values (r values were 0.916 and 0.608,all P＜0.01).Area under ROC curve of LS value for differentiating hepatic fibrosis stage were 0.938 to 0.989,while the areas of T1ρ were 0.771 to 0.954.Conclusion MR elastography is an accurate technique for quantitatively staging hepatic fibrosis and superior to T1ρ imaging.

Objective To analyze the pathological and clinical characteristics of patients with idiopathic IgA nephropathy accompanied by vasculitic/crescentic lesion (IgA-V/C). Methods Data of 222 patients diagnosed as idiopathic IgA nephropathy by renal biopsy, among them 87 cases with vasculitic/crescenlic (V/C)lesion, from our department in 2004 were analyzed retrospectively. Clinical and pathological data from patients with IgA-V/C were compared to those of non-IgA-V/C patients and of lupus nephritis (LN) patients with V/C lesion. Results Vasculitic/crescentic lesion was found in 39.19% (87/222) patients with idiopathic IgA nephropathy.And about( 14.08?12.75)% of the glomeruli was affected. It should be taken into account that there was no significant differences of clinical manifestations including blood pressure, urinary protein excretion between IgA-V/C group and non-IgA-V/C group .except serum creatinine(Scr)level which was significantly higher in IgA-V/C group. In addition, only 37.9% of IgA-V/C patients presented high Scr level,thus the lesion of V/C in idiopathic IgA nephropathy was easily overlooked. Patients with idiopathic IgA nephropathy were found to have higher percentage of glomerular sclerosis (64.86% vs 40.00%) and ratio of sclerostic glomeruli to total glomeruli [ (26.98 ?24.68)% vs (16.10 ?18.80)% ]as compared to LN group, which further predicated the progressing characteristics of IgA nephropathy. Conclusions Vasculitic/crescentic lesion is a quite common finding in idiopathic IgA nephropathy and often associated with no dramatically symptoms. It is possible for vasculitic/crescentic lesion to induce unmarked lose of nephron slowly and continually, so as to accelerate IgA nephropathy progression to end-stage renal failure.

Objective To evaluate the feasibility and effectiveness of combination chemotherapy with etoposide and cisplatin(EP)regimen on the patients with high-risk,chemorefractory and recurrent gestational trophoblastic neoplasia(GTN).Methods Thirty-nine patients with gestational trophoblastic tumors were analyzed retrospectively,25 of 39 patients were of high-risk,9 patients were chemorefractory and 5 patients were recurrent.All 39 patients were administrated with EP regimen,and 10 patients were assisted with surgery.All the patients were followed up.Clinical response,toxicity,the occurrence of secondary tumors of all patients,and the fertility of 30 patients whose fertility function was preserved were investigated. Results Thirty-nine GTN patients underwent a total of 221 cycles of the EP regimen.The average number of courses for each patient was 5.7.The total complete remission rate of the regimen was 74%(29/39). Twenty-five patients with high-risk GTN received a total of 139 cycles and the average number of courses was 5.6.Nineteen patients achieved complete remission and 6 patients showed drug-resistant.The complete remission rate of the high-risk group was 76%(19/25).Nine patients with chemorefractory GTN obtained a total of 55 cycles and the average number of courses was 6.1.Six patients achieved complete remission and 3 patients showed drug-resistant again.The complete remission rate of the chemorefractory group was 6/9. Five patients with recurrent GTN received 27 cycles and the average number of courses was 5.4.Four patients achieved complete remission,1 patient showed drug-resistance and died.Bone marrow toxicity, gastrointestinal reaction and alopecia were the main side effects of the EP regimen,but the bone marrow toxicity was slight and no grade Ⅳ side effect occurred.No fatal effect was found.Eight of 30 patients whose fertility faction was preserved had become pregnant after recovery,with a total of 8 pregnancies.Among them,2 were terminated by induced abortion,and 6 underwent normal term delivery and gained 6 infants who had no congenital malformation.All the 6 children had normal growth and development after childbirth. None of the women developed secondary tumors.Conclusion The EP regimen is effective and safe for the treatment of high-risk,chemorefractory and recurrent GTN.

AIMTo verify the hypothesis that repeated body position change training can improve human head-down tilt (HDT) tolerance.

METHODSSix young healthy subjects were trained with repeated position change for 9 times and 11 days according to protocol of alternative head-down and head-up tilts, each time of training lasted for about 35 min. Their HDT tolerance (- 30 degrees/30 min) were determined before and after training.

RESULTS(1) Compared with the data before training, subjects' symptom scores during HDT test after training decreased significantly (6.00 +/- 3.79 vs 1.00 +/- 0.63, P < 0.05), magnitude of the decreased heart rate increased significantly (-0.6 +/- 2.5 vs -4.4 +/- 3.6, P < 0.01). (2) Before training, blood flow volume of internal jugular vein (IJV) during HDT decreased significantly and that of internal carotid artery (ICA) increased significantly at the beginning period of HDT compared with pre-HDT (P < 0.01), while blood flow volume of the common carotid artery (CCA) presented increasing trend. After training, there was no significant difference in blood flow volume of IJV between during HDT and pre-HDT, that of ICA and CCA presented decreasing trend in the final period of HDT compared with Pre-HDT.

CONCLUSIONRepeated body position change training can improve human head-down tilt tolerance. And its main causation is that headward shift of blood volume is restrained to some extend during HDT after training.

Adaptation, Physiological ; physiology ; Adolescent ; Astronauts ; Cardiovascular Physiological Phenomena ; Head-Down Tilt ; Humans ; Male ; Posture ; physiology ; Weightlessness Simulation ; Young Adult

OBJECTIVETo observe the effect of three Chinese medical formulae (Zhifei Mixture I , Zhfei Mixture II, and Zhifei Mixture II) on main and secondary symptoms and signs of children with Totally 70 mycoplasma pneumonia in treating three types of children mycoplasma pneumonia.

METHODSchildren with mycoplasma pneumonia were assigned to the control group (38 cases) and the treatment group (32 case). All patients were intravenously injected with Azithromycin and took Ambroxol Hydrochloride and Clenbuterol Hydrochloride Oral Solution. Those in the treatment group additionally took Zhifei Mixture I , Zhfei Mixture II, and Zhifei Mixture Ill by syndrome typing. Their main and secondary symptoms and signs were observed before and after treatment (main symptoms and signs covered fever, cough, abundant sputum, short breath, and anoxia; secondary symptoms and signs covered aversion to cold, heart rate, facial complexion, spirit, appetite, and sweating).

RESULTSSeven patients were lost in this study. Compared with before treatment in the same group, scores for main and secondary symptoms and signs decreased in the treatment group (P <0.01). The therapeutic effect on fever and cough was obviously better in the control group (P <0.01). The main and secondary symptoms and signs were more obviously improved in the treatment group than in the control group (P <0.01). Commpared with the control group, scores for main and secondary symptoms and signs decreased more in the treatment group (P <0.01). Patients' main and secondary symptoms and signs were more obviously improved (P <0.05).

CONCLUSIONSZhifei Mixture combined Western drugs could significantly improve main and secondary symptoms and signs of mycoplasma pneumonia children patients. Its efficacy was superior to that of using Western medicine alone.

Ambroxol ; administration & dosage ; therapeutic use ; Anti-Bacterial Agents ; administration & dosage ; therapeutic use ; Azithromycin ; administration & dosage ; therapeutic use ; Bronchodilator Agents ; administration & dosage ; therapeutic use ; Child ; Clenbuterol ; administration & dosage ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Expectorants ; administration & dosage ; therapeutic use ; Fever ; Humans ; Pneumonia, Mycoplasma ; drug therapy ; Syndrome

OBJECTIVETo adapt the candidate strains of hemorrhagic fever with renal syndrome (HFRS) purified vaccine to Vero cells and to study their antigenicity and immunogenicity.

METHODSThe viral strains H8207 (Hantaan virus, HTN) and Y86013 (Seoul virus, SEO) were continuously propagated in Vero cell by the terminal dilution method and studied the characteristics of virus multiplication, viral titers and the amounts of virus antigen after serial passages. Three batches of crude monovalent inactivated vaccine were developed using the different passages of these 2 viral strains.

RESULTSThe strains H8207 and Y86013 adapted to Vero cells and stably grew on the cells with high titers. Rabbits immunized with the crude vaccines of H8207 and Y86013 showed 100% sero-conversion and the neutralizing antibody titers of the rabbit immune sera reached 1?10 at 4 weeks after 2 times of immunization.

CONCLUSIONSThe results suggest that these 2 candidate strains had adapted to Vero cells, possessed high titers and good immunogenicity and be feasible to prepare the HFRS purified vaccine in Vero cells.

Animals ; Antibodies, Viral ; blood ; Cercopithecus aethiops ; Hantaan virus ; growth & development ; immunology ; Hemorrhagic Fever with Renal Syndrome ; prevention & control ; Mice ; Neutralization Tests ; Rabbits ; Seoul virus ; growth & development ; immunology ; Vaccination ; Vaccines, Inactivated ; immunology ; Vero Cells ; Viral Vaccines ; biosynthesis ; immunology

OBJECTIVETo investigate the effect of sphingosine kinase 1 (SphK1) on the proliferation, apoptosis, migration and invasion of colon cancer TH-29 cells and to explore its molecular mechanisms.

METHODSPhorbol 12-myristate 13-acetate (PMA) was used to induce the activity of SphK1 and N, N-dimethylsphingosine (DMS) was used to suppress the activity of SphK1. Cell prolieration and apoptosis were detected by MTT assay and flow cytometry, respectively. The migration and invasion capabilities of the cells were assessed in Transwell chambers. The activity of SphK1 was assayed by autoradiography. Western blot was used to evaluate the protein expression of SphK1, p38, phosphorylated p38 (p-p38) and SAPK/JNK.

RESULTSPMA and DMS were able to induce and suppress the activity and protein expression of SphK1 in a time-dependent manner, respectively. PMA enhanced and DMS suppressed the cell viability in a time- and dose-dependent manner. Being treated with 100 nmol/L PMA or 50 µmol/L DMS for 0, 6, 12, 24 h, the cell apoptosis rates of PMA group were (9.35 ± 0.84)%, (7.61 ± 0.48)%, (5.53 ± 0.76)% and (0.56 ± 0.33)%, contrastly, that of DMS group were (9.18 ± 0.94)%, (12.06 ± 1.41)%, (19.80 ± 2.36)% and (31.85 ± 3.60)%, respectively. Compared with the control group, the cell migration and invasion capabilities of the PMA group were significantly enhanced, and that of the DMS group were significantly suppressed. The migration cell number of control, PMA and DMS groups were 68.75 ± 6.15, 109.33 ± 11.63 and 10.83 ± 2.48, the invasion cell number of control, PMA and DMS groups were 55.42 ± 4.50, 90.58 ± 7.06 and 9.58 ± 2.39, respectively. With the elevating activity and expression of SphK1, the protein expressions of p38, p-p38 and SAPK/JNK were strikingly suppressed. On the contrary, after treating with DMS the protein expressions of p38, p-p38 and SAPK/JNK were enhanced.

CONCLUSIONSSphK1 potently enhances the prolieration, migration and invasion of colon cancer HT-29 cells, meanwhile suppresses the cell apoptosis. The suppressing of the p38 and SAPK/JNK signalling pathways may be one of its molecular mechanisms.

Apoptosis ; drug effects ; Carcinogens ; administration & dosage ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Enzyme Inhibitors ; administration & dosage ; pharmacology ; HT29 Cells ; Humans ; MAP Kinase Kinase 4 ; metabolism ; Neoplasm Invasiveness ; Phosphorylation ; Phosphotransferases (Alcohol Group Acceptor) ; metabolism ; physiology ; Sphingosine ; administration & dosage ; analogs & derivatives ; pharmacology ; Tetradecanoylphorbol Acetate ; administration & dosage ; analogs & derivatives ; pharmacology ; Time Factors ; p38 Mitogen-Activated Protein Kinases ; metabolism

To combine the fibrinolytic with anticoagulant activities for therapy of thrombotic deseases, a fusion protein made of tissue-type plasminogen activator (t-PA) and hirudin was constructed and expressed in chia pastoris. To improve thrombolytic properties of t-PA and reduce bleeding side effect of hirudin, FXa-recognition sequence was introduced between t-PA and hirudin molecules.The anticoagulant activity of hirudin can be target-released through cleavage of FXa at thrombus site. t-PA gene and hirudin gene with FXa-recognition sequence at its 5'-terminal were obtained by RT-PCR and PCR respectively. The fusion protein gene was cloned into plasmid pIC9K and electroporated into the genome of Pichia pastoris GS115. The expression of fusion protein was induced by methanol in shaking flask and secreted into the culture medium. Two forms of the fusion protein, single-chain and double-chain linked by a disulfide bond (due to the cleveage of t-PA at Arg275-Ile276), were obtained. The intact fusion protein retained the fibrinolytic activity but lacked any anticoagulant activity. After cleavage by FXa, the fusion protein liberated intact free hirudin to exert its anticoagulant activity. So, the fusion protein is a bifunctional molecule having good prospect to develop into a new targeted therapeutic agent with reduced bleeding side effect for thrombotic diseases.
Animals ; Cloning, Molecular ; Electroporation ; Hirudins ; biosynthesis ; genetics ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Plasminogen Activator ; biosynthesis ; genetics

Traditional T vector cloning method requires onerous procedures for identifying recombinant, and directional cloning was impossible. In order to overcome these problems, we have devised a directional T vector pETG based on pET-23a(+). For gene cloning, 7 bp partial LacO sequence was introduced into DNA fragment to reconstitute a full length LacO with Bfu I digested T vector. After transformation, blue colonies were selected on LB plate supplemented with X-gal. Restriction enzyme digestion and PCR identification showed that all blue colonies contained the directionally inserted recombinants and the recombinant efficiency was nearly 100%. We have successfully cloned 103 genes from human liver cDNA; in the study complicated procedures for screening of recombinant were not required. Eight pETG clones were picked for protein expression, and all the clones successfully produced corresponding proteins. We demonstrated that the directional T vector was successfully constructed, and it was very suitable for gene cloning and expression.
Cloning, Molecular ; methods ; DNA, Complementary ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; genetics ; Genetic Vectors ; genetics ; Humans ; Liver ; chemistry ; Polymerase Chain Reaction ; methods ; Recombinant Proteins ; biosynthesis ; genetics