Fosmidomycin (100 micromol x L(-1)) which is the effective inhibitor of DXR, key enzyme in terpenoid MEP pathway, was used to treat with hairy roots of Salvia miltiorrhiza. The treated roots were harvested at 2, 4, 6, 8, 10, 16 and 21 d, mRNA level of SmDXR and tanshinone content in treated and negative control groups were detected. Results found that, after treated with fosmidomycin, color of S. miltiorrhiza hairy roots grew pale gradually comparing with controls; mRNA level of SmDXR in hairy roots varied as a shape of parabolic and the highest value achieved at the sixth day after treatment, then it decreased gradually; Content of four kinds of tanshinones were detected. Among of the four kinds of tanshinones, Tanshinone I content changed relatively little, while content of dihydrotanshinone I, cryptotanshinone and tanshinone II (A) decreased gradually in 21 days. The content of total tanshinones in NC groups was 5, 63 times more than FOS-treated roots in the 21th day. The previous results showed that SmDXR played an important role in the accumulation of tanshinone content in MEP pathway. Once the mRNA level of SmDXR was suppressed, the accumulation of secondary metabolites will be significantly affected.
Aldose-Ketose Isomerases ; genetics ; Diterpenes, Abietane ; metabolism ; Fosfomycin ; analogs & derivatives ; pharmacology ; Gene Expression Regulation, Plant ; drug effects ; Plant Roots ; drug effects ; growth & development ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Salvia miltiorrhiza ; drug effects ; genetics ; growth & development ; metabolism ; Time Factors

Objective To investigate the MR features of pituitary abscess.Methods The MR features of 14 cases of pituitary abscess proved by surgical pathology and clinical treatments were analyzed retrospectively.Results Pre-contrast MR showed hypointense heterogeneous intensity on T_1 WI in 12 cases and iso-hyperintense on T_1 WI in 2 cases,hyperintense on T_2 WI in all cases.Post-gadolinium MR showed the ring-like enhancement around the uneven edge of abscess and the surrounding enhanced meninges connecting to the focus.The normal pituitary could not be identified in all 14 cases.The MR specific findings include the fluid-fluid level,nodule on the edge and the enhanced patchy shadow.Conclusions The pituitary abscess has specific findings on MR examination,which can be used to combine with clinical symptoms to achieve the diagnosis before operation,so that the cases could be treated with antibiotic without operation.

The chemical investigation on Ganoderma philippii led to the isolation of sixteen compounds by silica gel and Sephadex LH-20 column chromatography. On the basis of spectroscopic data analyses, their structures were elucidated as 2, 5-dihydroxyacetophenone (1), methyl gentisate (2), (S) -dimethyl malate (3), muurola-4, 10 (14) -dien-11beta-ol (4), dihydroepicubenol (5), 5-hydroxymethylfuran carboxaldehyde (6), ergosta-7, 22E-dien-3beta-ol (7), ergosta-7, 22E-dien-3-one (8), ergosta-7, 22E-diene-2beta, 3alpha, 9alpha-triol (9), 6/beta-methoxyergo-sta-7, 22E-dien-3beta, 5alpha-diol (10), ergosta-4, 6, 8(14), 22E-tetraen-3-one (11), ergosta4, 6, 8-(14), 22E-etetraen-3beta-ol (12), 5alpha, 8alpha-epidioxy-ergosta-6, 22E-dien-3beta-ol (13), 7alpha-methoxy-5alpha, 6alpha-epoxyergosta-8-(14), 22E-dien-3beta-ol (14), ergosta-8, 22E-diene-3beta, 5alpha, 6beta, 7alpha-tetraol (15), and ergosta-5, 23-dien-3beta-ol, acetate (16). All the compounds were obtained from this fungus for the first time, and compounds 4 and 5 were isolated from the Ganoderma genus for the first time.
Ganoderma ; chemistry ; Medicine, Chinese Traditional ; Organic Chemicals ; analysis ; isolation & purification

BACKGROUNDHIV-1 infected and immune-activated macrophages and microglia secrete neurotoxins, such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), which play major role in the neuronal death. It has been shown that different HIV-1 variants have varying abilities to elicit secretion of TNF-α by peripheral blood mononuclear cell (PBMC); however, whether the difference of gp120 gene could affect the secretion of TNF-α and IL-1β by glial cells is unknown. The aim of this study was to explore the association between gene diversity and induction of neurotoxic cytokines.

METHODSIn this study, we constructed retroviral vectors MSCV-IRES-GFP/gp120 using HIV-1 gp120 genes isolated from four different tissues of one patient who died of AIDS dementia complex (ADC). Recombinant retroviruses produced by cotransfection of MSCV-IRES-GFP/gp120, pCMV-VSV-G and pUMVC into 293T cells were collected and added into U87 glial cells. Concentrations of TNF-α and IL-1β secreted by transduced U87 cells were assayed with ELISA separately.

RESULTSThe four HIV-1 gp120 were in the different branch of the neighbor-joining tree. Compared to the pMIG retrovirus (gp120-negative) or U87 cells, all the gp120-positive recombinant retroviruses induced more TNF-α (P < 0.01) and IL-1β (P < 0.01). In addition, compared with the L/MIG retrovirus, all the three brain gp120-positive recombinant retroviruses induced less TNF-α (P < 0.01) and IL-1β (P < 0.01).

CONCLUSIONSHIV-1 gp120 could induce U87 cells secret more TNF-α and IL-1β again. The more important is that difference of HIV-1 gp120, especially cell-tropism may account for the different ability in eliciting secretion of TNF-α and IL-1β, which might supply a novel idea helping understand the pathogenesis of ADC.

AIDS Dementia Complex ; metabolism ; virology ; Cell Line ; Cell Line, Tumor ; Enzyme-Linked Immunosorbent Assay ; HIV Envelope Protein gp120 ; genetics ; metabolism ; Humans ; Interleukin-1beta ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

In this study, the inhibitory effect of human umbilical cord-derived mesenchymal stem cells (hUCMSC) on interleukin-17 (IL-17) production in peripheral blood T cells from patients with spondyloarthritis (SpA) were investigated, in order to explore the therapeutic potential of hUCMSC in the SpA. Peripheral blood mononuclear cells (PBMNC) were isolated from patients with SpA (n = 12) and healthy subjects (n = 6). PBMNC were cultured in vitro with hUCMSC or alone. The expression of IL-17 in CD4(+) T cells or γ/δ T cells were determined in each subject group by flow cytometry. IL-17 concentrations in PBMNC culture supernatants were measured by ELISA. The results indicated that the proportion of IL-17-producing CD4(+) T cells and IL-17-producing γ/δ T cells of SpA patients were 4.5 folds and 5 folds of healthy controls [CD3(+)CD4(+)IL-17(+) cells (3.42 ± 0.82)% vs (0.75 ± 0.25)%, P < 0.01; CD3(+)γδTCR(+)IL-17(+) cells (0.30 ± 0.10)% vs (0.06 ± 0.02)%, P < 0.01]. After co-culture of PBMNC in patients with hUCMSC, the increased proportions of CD3(+)CD4(+)IL-17(+) cells and CD3(+)γδTCR(+)IL-17(+) cells in SpA patients were inhibited significantly by hUCMSC [CD3(+)CD4(+)IL-17(+) cells (3.42 ± 0.82)% vs (1.81 ± 0.59)% (P < 0.01); CD3(+)γδTCR(+)IL-17(+) cells (0.30 ± 0.10)% vs (0.16 ± 0.06)% (P < 0.01]. In response to phytohemagglutinin (PHA, 1 µg/ml), PBMNC from SpA patients secreted more IL-17 than that from healthy control [(573.95 ± 171.68) pg/ml vs (115.53 ± 40.41) pg/ml (P < 0.01)]. In the presence of hUCMSC, PBMNC of SpA patients produced less amount of IL-17 [(573.95 ± 171.68) pg/ml vs (443.20 ± 147.94) pg/ml, (P < 0.01)]. It is concluded that the IL-17 production in peripheral blood T cells from SpA patients can be inhibited by hUCMSC, which have therapeutic potential for SpA.
Humans ; Interleukin-17 ; metabolism ; Leukocytes, Mononuclear ; cytology ; Lymphocyte Count ; Mesenchymal Stromal Cells ; Spondylarthritis ; blood ; metabolism ; therapy ; T-Lymphocytes ; metabolism ; Umbilical Cord ; cytology

OBJECTIVETo explore the molecular mechanism of Qiangxin Fumai granule (QFG, an effective Chinese composite drug) in preventing and treating sick sinus syndrome (SSS).

METHODRabbit model of sinoatrial ischemia/reperfusion was established by occluding and loosening the root of right coronary artery. Effect of QFG on cell apoptosis was observed by TUNEL method, and its effect on apoptotic related gene Bax, Bcl-2 and Fas-L gene protein expression was observed by immunohistochemical method. Average light density values of the expression of Bax, Bcl-2 and Fas-L of SAN cells was determined by Imagepro Plus image analysis system.

RESULTSinoatrial injury induced by ischemia/reperfusion could cause evident sinoatrial cell apoptosis, enhance Fas-L gene protein expression and obviously enhance Bax gene protein expression, reduce Bcl-2/Bax ratio. QFG could significantly down-regulate Fas-L and Bax gene protein expression, up-regulate Bcl-2/Bax ratio, significantly inhibit and block the sinoatrial cell apoptosis.

CONCLUSIONTo inhibit and block the event of cell apoptosis through regulating Bax, Bcl-2 and Fas-L gene protein expression in sinoatrial node after ischemia/reperfusion might be one of the mechanisms of QFG in preventing and treating sinoatrial ischemia/reperfusion injury of SSS.

Animals ; Apoptosis ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fas Ligand Protein ; metabolism ; Immunohistochemistry ; In Situ Nick-End Labeling ; In Vitro Techniques ; Microscopy, Fluorescence ; Muscle Cells ; cytology ; drug effects ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rabbits ; Random Allocation ; bcl-2-Associated X Protein ; metabolism

Aqueous dispersion and stability of Fe3O4 nanoparticles remain an issue unresolved since aggregation of naked iron nanoparticles in water. In this study, we successfully synthesized different Fe3O4 super-paramagnetic nanoparticles which were modified by three kinds of materials [DSPE-MPEG2000, TiO2 and poly acrylic acid (PAA)] and further detected their characteristics. Transmission electron microscopy (TEM) clearly showed sizes and morphology of the four kinds of nanoparticles. X-ray diffraction (XRD) proved successfully coating of the three kinds of nanoparticles and their structures were maintained. Vibrating sample magnetometer (VSM) verified that their magnetic properties fitted for the super-paramagnetic function. More importantly, the particle size analysis indicated that Fe3O4@PAA had a better size distribution, biocompatibility, stability and dispersion than the other two kinds of nanoparticles. In addition, using CNE2 cells as a model, we found that all nanoparticles were nontoxic. Taken together, our data suggest that Fe3O4@PAA nanoaparticles are superior in the application of biomedical field among the four kinds of Fe3O4 nanoparticles in the future.

Objective To study the influences of Yang-warming, Blood-activating, and Toxin-removing Recipe(YBTR)-containing serum sampled under different conditions on the proliferation of human umbilical vein endothelial cells(HUVECs). Methods HUVECs were cultured in vitro for the experiment. Pharmacological serum for the experiment was prepared as follows: Thirty-two SD rats (male in half) were randomly divided into 4 groups, namely normal saline group and low-, middle-, and high-dose YBTR groups (at the intragastric dosage of 10.35, 31.05, 93.15 g·kg-1 respectively, twice a day). The pharmacological serum was taken from one female rat and one male rat in various groups at 4 time points, i.e. at hour 1, 2 after first intragastric administration on the fourth feeding day, and at hour 1, 2 after first intragastric administration on the sixth feeding day(abbreviated as D3H1, D3H2, D5H1, D5H2 respectively). The effects of YBTR-containing serum on the proliferation of HUVECs were observed by CCK-8 assay method. Results The difference of proliferation-inhibition rate of HUVECs was statistically significant after treated with YBTR-containing serum prepared from rats of different genders at different time(Pgender=0.000<0.01, Ptime=0.000<0.01). The difference of interaction of time and gender was also significant (Ptime × gender=0.001<0.01), and the effect at D3H1 and D5H1 varied with the gender (PD3H2×gender = 0.000 < 0.01, PD5H1×gender = 0.002 < 0.01). The inhibitory action of YBTR-containing serum became stronger with the increase of the dosage of serum of female rat at D3H2, and the inter-group difference was statistically significant (P < 0.05), the effect showing concentration-dependent tendency. The inhibition of HUVECs proliferation reached the peak after treated by various doses of YBTR-containing serum from the female rat at D3H2, while the inhibition arrived to the peak after treated by low- and middle-dose YBTR-containing serum from the male rat at D5H1, and the inhibition arrived to the peak in the group of high-dose YBTR-containing serum from the male rat at D3H1. Conclusion The inhibitory action of YBTR-containing serum on the proliferation of HUVECs was stronger when the serum was taken from the female rat at D3H2.

OBJECTIVETo investigate the risk factors for postoperative pulmonary complications (PPC) after gastroduodenal operation.

METHODSFrom December 1999 to December 2003, clinical data of 508 patients undergoing gastroduodenal operation were analyzed retrospectively. Risk factors for PPC were screened.

RESULTSThe complication rate of PPC was 25.8% (131/508). Multivariate logistic regression analysis revealed that age (OR=1.052), history of respiratory diseases (OR=2.915), serum albumin level (OR=0.995), length of intratracheal intubation (OR=1.005), length of nasogastric intubation (OR=1.059) and length of postoperative mechanical ventilation (OR=1.367) were risk factors for PPC.

CONCLUSIONPatients with old age, lower serum albumin level, intraoperative or postoperative nasogastric intubation, intratracheal intubation or long-term mechanical ventilation were more prone to develop PPC.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Digestive System Surgical Procedures ; adverse effects ; Duodenum ; surgery ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Pneumonia ; epidemiology ; Postoperative Complications ; epidemiology ; Respiratory Distress Syndrome, Adult ; epidemiology ; Retrospective Studies ; Risk Factors ; Stomach ; surgery ; Young Adult

OBJECTIVETo analyze the meiotic segregation results of the spermatozoa from male pericentric inversion carriers by fluorescence in-situ hybridization (FISH).

METHODSUsing chemical depolymerization and multicolor FISH, we analyzed the meiotic segregation results of the spermatozoa from 4 male pericentric inversion carriers.

RESULTSOf the 4 males studied, 46,XY,inv(9) (p11q12) was found in 2, 46,XY,inv(9) (p11q13) in 1 and 46,XY,inv(6) (p22q24) in the other; the lengths of the inverted segments represented 16.0, 16.0, 21.0 and 76.0% of the size of the whole chromosome involved; and the frequencies of recombinant sperm were 0.2, 0.4, 0.3 and 43.9%, del(p)/dup(q) accounting for 22.4% and del(q)/dup(p) 21.5%, respectively.

CONCLUSIONMales with pericentric inversion may produce spermatozoa with recombinant chromosomes and the rate of recombination varies principally according to the size proportion to the whole chromosome involved. The results of FISH analysis of chromosomal unbalanced spermatozoa can provide accurate personalized information on the genetic risk of fertility.

Adult ; Chromosome Inversion ; genetics ; Chromosomes, Human, Pair 9 ; genetics ; Heterozygote ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infertility, Male ; genetics ; Male ; Meiosis ; Spermatozoa