Ten compounds, including seven sesquiterpenes, two phenols and one phenylpropanoid, were isolated from the roots of Illicium majus by means of silica gel, ODS, Sephadex LH-20, and preparative HPLC. On analysis of MS and NMR spectroscopic data , their structures were established as cycloparviflorolide (1), cycloparvifloralone (2), tashironin (3), tashironin A (4), anislactone A(5), anislactone B (6), pseudomajucin (7), syringaldehyde (8), methyl-4-hydroxy-3, 5-dimethoxybenzoate (9), and (E)-3-methoxy-4,5-methylenedioxycinnamic alchol (10), respectively. Compounds 1-4 and 8-10 were first isolated from this plant. In the in vitro assays, at a concentration of 1.0 x 10(-5) mol x L(-1), compounds 5 and 6 were active against LPS induced NO production in microglia with a inhibition rate of 75.31% and 53.7%, respectively.
Drugs, Chinese Herbal ; analysis ; chemistry ; Illicium ; chemistry ; Organic Chemicals ; analysis ; chemistry ; Plant Roots ; chemistry

OBJECTIVEAnalyzing the relationships between peripheral blood CD4+ CD25hi regulatory T (Treg) cells and peripheral blood immune status or plasma HIV-lviral load in HIV-infected individuals,so as to determine whether Treg were related to the progression of HIV-infected disease.

METHODS116 HIV-infected patients in different stages and 21 healthy control individuals were included in this study. The CD4+ and CD8+ T cell counts were determined by a standard 4-color flow cytometry technique. The Treg cells were examined with 3-color immune staining flow cytometry. The plasma HIV-1 viral load was detected by real time PCR.

RESULTSThe frequencies of Treg cells decreased in HIV-infected individuals with high CD4+ T cell counts( > 300/microl) compared with normal controls. With the progression of disease the frequencies of Treg cells were raised gradually, until were increased in HIV-infected individuals with low levels of CD4+ T cell counts ( < 100/microl). In addition, the frequencies of Treg cells were inversely related to CD4+ T cell counts and CD4+ /CD8+ ratio, data showed a statistically significant (respectively, r = -0.564, P < 0.001; r = -0.377, P < 0.001). Furthermore, the proportions of Treg cells were closely related to plasma HIV-1 RNA viral load (r = 0.514, P < 0.001).

CONCLUSIONCD4 CD25hi Treg cells should be a kind of important cells participating the immunopathogenesis of AIDS. It may play different roles in different stages of HIV-infected disease. The exact mechanism of Treg cells in the progression of the HIV-infected disease needs to be investigate further.

Adult ; Case-Control Studies ; Cells, Cultured ; Disease Progression ; Female ; HIV Infections ; immunology ; pathology ; virology ; HIV-1 ; genetics ; immunology ; isolation & purification ; Humans ; Interleukin-2 Receptor alpha Subunit ; immunology ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology ; Viral Load

OBJECTIVETo provide DNA molecular marker for identification of Nelumbo nucifera by exploring the differences of nrDNA-ITS sequence of N. nucifera originated from different habitats.

METHODTo compare nrDNA-ITS base sequence using specific PCR-ITS.

RESULTThe completed sequence of ITS and 5.8 S rDNA, and the partial sequences of 18S rDNA and 26S rDNA, totally 750 bp, from N. nucifera were obtained. The differences among N. nucifera from different habitats and from different cultivars were found.

CONCLUSIONThe method can be used to identify N. nucifera among different species and to distinguish their fakes. It provided the basis for identifying N. nucifera from different geographical regions by comparison of their ITS sequences.

Base Sequence ; China ; DNA, Plant ; chemistry ; genetics ; metabolism ; DNA, Ribosomal Spacer ; classification ; genetics ; Deoxyribonuclease EcoRI ; metabolism ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Drug Contamination ; prevention & control ; Geography ; Nelumbo ; classification ; genetics ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction ; RNA, Ribosomal ; genetics ; RNA, Ribosomal, 18S ; genetics ; RNA, Ribosomal, 5.8S ; genetics ; Sequence Analysis, DNA ; Species Specificity

Objective: To facilitate the functional analysis of chromosomal genes and their products, the recombineering technique to epitope tagging of chromosomal genes of Y. pestis was adapted. Methods: The epitope tag was generated by primer annealing and then fused with resistance gene by fusion PCR. The epitope-resistance cassette was inserted into pBluecript, resulted in the template plasmid, pBS-MH. The tagging cassette for rpoS was obtained by PCR amplification from pBS-MH with primers containing homology specific to the target gene. PCR products were transformed into recombination competent cells and recombinants were selected. PCR and DNA sequencing were used to confirm the correct tagging event. The expression of the tagged protein was detected with Western blot by using monoclonal antibody to the epitope. Results: The template plasmid containing fusion of epitope and resistance gene was successfully constructed. The sigma factor gene, rpoS, was tagged with a myc-his tag at the COOH terminus. Expression of the tagged rpoS was successfully detected indirectly by the antibody against His tag. Conclusion: The chromosomal gene tagging by recombineering technique represents a powerful tool in the functional study of bacterial genes and their products.

OBJECTIVETo study the chemical constituents of Ginseng Sini Tang.

METHODThe constituents were identified by physico-chemical properties and spectral analysis.

RESULTThe 12 compounds were identified as ginsenoside-Rb1,-Rb2,-Rb3,-Rc,-Rd,-Re,-Rg1,Rg2,Rg3,Rf,Ra1,Ra2. The 10 compounds were identified as benzoylmesaconitine(BM), benzoylaconitine(BA), benzoylhypaconitine(BH), neoline (NL), fuziline (FL), 14-ethyl-talatisamine14-acetyl-talatisamine (AT), 14-benzoylhypaconine-8-linoleate (HAL),14-benzoyldeoxyaconine-8-oleate(HAO), 14-benzoylhypaconine-8-palmitate(HAP), talatisamine(TS).

CONCLUSIONAll these compounds were obtained from Ginseng Sini Tang for first times.

Alkaloids ; isolation & purification ; pharmacology ; Animals ; Depression, Chemical ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Ginsenosides ; isolation & purification ; pharmacology ; Myocardial Contraction ; drug effects ; Panax ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVETo observe the outcome of percutaneous balloon mitral valvuloplasty (PBMV) in patients with rheumatic mitral valve stenosis.

METHODSFrom April 1992 to November 2008, 1768 patients underwent PBMV in our hospital.Clinical and echocardiographic follow up data were analyzed in 426 patients from April 1992 to August 1998. Left atrial pressure and the mitral valve gradient (MVG) were measured before and immediately after PBMV in all patients.

RESULTSPBMV was successful in 1748 out of 1768 patients (98.86%). Left atrial pressure decreased from (38 +/- 7) mm Hg (1 mm Hg = 0.133 kPa) to (12 +/- 4) mm Hg (P < 0.001), MVG decreased from (28 +/- 6) mm Hg to (8 +/- 3) mm Hg (P < 0.001) and the area of the mitral valve increased from (0.98 +/- 0.26) cm(2) to (1.97 +/- 0.39) cm(2) (P < 0.001) post PBMV. The main complications included death (n = 2), acute pericardial effusion (n = 1), severe mitral regurgitation (n = 12), cerebral embolism (n = 2) and pulmonary edema (n = 1). Ten years follow up was finished in 426 patients and 288 patients (67.6%) were still in NYHA class Ior II without mitral valve replace operation or repeated PBMV, restenosis was evidenced in 140 patients (33.3%) and 31 patients dead (7.5%).

CONCLUSIONPBMV was an effective therapy option for patients with rheumatic mitral valve stenosis.

Catheterization ; adverse effects ; Echocardiography ; Follow-Up Studies ; Humans ; Mitral Valve Stenosis ; therapy ; Rheumatic Heart Disease ; therapy ; Treatment Outcome

Carrier State ; epidemiology ; Child ; China ; epidemiology ; Female ; Humans ; Male ; Streptococcal Infections ; epidemiology ; Streptococcus pneumoniae

Objective To explore and compare the relevant regional anatomies as they relate the fron- tolateral keyhole approach under microscopy and neuroendoscopy for operations in anterior cranial base and sellar region.Methods Fifteen silieone-injected cadaveric heads were dissected to reveal and compare the extent of expesure through the transfrontolateral keyhole approach under neuroendoscopy and microscopy. Results Portions in the areas of olfactory groove,sellar region and sylvian tissure were blind under micro- scope.Endoscope could allow observation of areas considered blind under the microscope.It could increase light intensity during the approach to objects,extend viewing angles,clear depiction of details in close-up po- sitions and inspect hidden structures.But images of endoscope were two dimensional,lack of view depth.Mi- croscopy and neuroendoscopy could help each other to recuperate deficiency.Conclusion Endoscope-assis- ted neuromicrosurgery is helpful,safe and minimally invasive to treat deepseated lesions in anterior cranial base,sellar region by transfrontolateral keyhole approach.

OBJECTIVETo construct the eukaryotic expression vector of human microdystrophin gene and observe its expression in rat mesenchymal stem cells (rMSCs) in vitro.

METHODSThe plasmid PBSK-MICRO containing human microdystrophin cDNA was digested by restriction endonuclease, and the resultant microdystrophin fragment was inserted into the NotI site of pcDNA3.1(+) to prepare the eukaryotic expression vector-pcDNA3.1(+)/ microdystrophin, which was identified by endonuclease digestion and sequencing. The recombinant plasmid was transfected into rMSCs via lipofectamine, and after G418 selection, the expression of microdystrophin was detected by RT-PCR and indirect immunofluorescence assay.

RESULTSMicrodystrophin gene fragment was correctly inserted into the plasmid pcDNA3.1(+), as conformed by sequencing and digestion with Not I and Hind III. The total mRNA of the transfected rMSCs was extracted and microdystrophin mRNA expression was found in the cells by RT-PCR. Indirect immunofluorescence assay for the protein expression of microdystrophin showed bright red fluorescence in the transfected rMSCs.

CONCLUSIONEukaryotic expression plasmid pcDNA3.1(+)/microdystrophin has been constructed successfully and microdystrophin can be expressed in transfected rMSCs in vitro, which may facilitate further research of Duchenne muscular dystrophy treatment by genetically modified allogeneic stem cell transplantation.

Animals ; Base Sequence ; Cells, Cultured ; Dystrophin ; biosynthesis ; genetics ; Fluorescent Antibody Technique, Indirect ; Gene Expression ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Molecular Sequence Data ; Peptide Fragments ; biosynthesis ; genetics ; Plasmids ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo evaluate the effect of PEGylated IL-11 mutein (PEG-mIL 11) with different dose or injection frequency on thrombocytopenia in myelosuppressed mice and to compare its effect with mIL-11, so as to provide reference data for clinical use.

METHODSMyelosuppressive model with thrombocyopenia was produced in BALB/c mice by whole bodyCo γ-ray irradiation in dose of administration 2.5 Gy followed by i.p. injections of carboplatin at 50 mg/kg. In study of injection frequency, 30 thrombocytopenic BALB/c mice were randomly divided into 5 groups: vehicle control group (once daily on d1, 4, 7), mIL-11 group [200 µg/(kg·d)×9 d], PEG-mIL 11 A group [111800 µg/(kg·d)×1 d (d 1)], PEG-mIL 11 B group 900 µg/(kg·d)×2 d (d 1,5), and PEG-mIL 11 C group [600 µg/(kg·d)×3 d (d 1,4,7)]. The route of administration is subcutaneous injection. The platelet counts were monitored in the subsequant 5 weeks. In study of dose administration, 100 thrombocytopenic BALB/c mice were randomly divided into 5 groups: vehicle control group (once daily on d1 and 5), mIL-11 group 200 µg/(kg·d)×9 d, and PEG-mIL 11 low-, mid-, and high-dose groups (200, 420 and 900 µg/(kg·d)×2 d, once a day on d1 and 5). The route of administration is subcutaneous injection. The platelet counts were monitored every 2-3 days in the subsequant 5 weeks, and CFU-Meg was determined on d 8 of the bone marrow cells collection.

RESULTSAfterCo irradition and carboplatin injection, Plt level decreased with time, and a >80% reduction was noted at nadir when comparing with baseline. In frequency of administration study, the platelet nadir of the 3 PEG-mIL 11 groups were significantly higher than that of vehicle control and mIL-11 groups (P<0.05), while no significant difference was noted among the 3 groups of different administration frequences; In dose level study, the reduction of Plt count at the nadir in the 3 PEG-mIL 11 groups was significantly less than that of vehicle control and mIL-11 groups (P<0.05). And a rapid recovery of Plt count was found in the PEG-mIL 11 groups with a dose-dependent increase of Plt count on d 10. A lower reduction and a rapider recovery of RBC was also found in the PEG-mIL 11 groups. No significant effect on WBC was found for all the treatment groups. An increase in CFU-Meg was observed in PEG-mIL 11 and mIL-11 groups, with higher CFU-Meg in PEG-mIL 11 groups.

CONCLUSIONA preventive effect of PEG-mIL 11 on thrombocytopenia in myelosuppressed mice has been confirmed. In comparison with mIL-11, a better effect of PEG-mIL 11 is obtained under lower dose frequency, indicating a better compliance of the treatment regimen, and providing a foundation for developing a long-acting preparation of rhIL-11.