Neural stem cells are a potential therapeutic source for cellular transplantation therapy in neurological diseases. The present paper was aimed to investigate whether neural stem cells could be obtained from the spinal cords of low temperature preserved abortuses. Fourteen weeks old abortuses were stored in a refrigerator at 4 degrees C without any additional treatments for 2, 6 and 12 h before use. The spinal cords were anatomized out and divided into cervical cords, thoracic cords and lumbar/sacral cords. Then the spinal cord segments were used for cell culture separately. Neural stem cells were isolated from the segments and cultured in bFGF, EGF and N2 supplement containing free-serum DMEM/F12 (1:1) medium. In order to examine the differentiation potential, the stem cells were induced to differentiate with 5% fetal bovine serum on poly-l-lysine substrate. Clonal culture was carried out to demonstrate that the isolated cells met the standard of stem cells. Indirect fluorescent immunocytochemistry was used to examine the expressions of neural stem cell marker (nestin), neuron marker (MAP2), astrocyte marker (GFAP) and cholinergic marker (ChAT). The stem cells in different cultures were compared. One-way analysis of variance and Kruskal-Wallis test were used for the statistical comparison. As a result, neural stem cells were obtained from all the spinal cord segments with different postmortem intervals. Both the cells on the surface and inside the neurospheres showed nestin immunoreactivity. Therefore, nearly all the cells that composed the neurospheres were nestin-positive undifferentiated cells. When the spheres were induced to differentiate, they could yield GFAP-positive astrocytes and MAP2-positive neurons including ChAT-positive cholinergic neurons. Primary neurospheres could be dissociated mechanically, expand in subcultures and maintain the differentiation potential. In clonal cultures, single cells from a single primary sphere could give rise to new neurospheres, which had the same differentiation potential as the primary spheres. The lumbar/sacral cord cultures gave rise to the most abundant primary neurospheres. When the preservation time of the fetus was prolonged to 12 h, the number of primary neurospheres decreased sharply. The clonal formation and phenotype capacity were similar in all cultures. In conclusion, spinal neural stem cells can be isolated from low temperature preserved abortuses and represent an alternative source for experimental and potential therapeutic purposes.
Cell Differentiation ; physiology ; Cell Separation ; Cells, Cultured ; Cryopreservation ; Culture Media, Serum-Free ; Fetus ; cytology ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Nestin ; metabolism ; Neural Stem Cells ; cytology ; Spinal Cord ; cytology

OBJECTIVETo explore the therapeutic effect of qi benefiting blood activating method (QB-BAM) on acute exacerbation of chronic obstructive pulmonary disease (AECOPD) patients with blood stasis syndrome (BSS) by observing its effect on plasma fibrinogen (Fg) and D-dimer (D-D) levels.

METHODSSixty AECOPD patients with BSS were randomly assigned to the treated group and the control group, 30 in each group. All patients received conventional therapy for AECOPD. Those in the treated group were additionally injected with Shengmai Injection and Tanshinone IIA Injection. Clinical efficacy and indices including levels of Fg, D-D, PaO2, and PaCO2 were measured and compared before and after treatment.

RESULTSThe effective rate was 93.3% (28/30 cases) in the treated group, higher than that of the control group [73.3% (22/30 cases) , P < 0.05]. There was no significant difference in all indices between the treated group and the control group before treatment (P >0.05). After treatment all indices were significantly improved in the two groups (P < 0.01). But in the treated group levels of Fg and D-D decreased more and levels of PaO2 increased more (P < 0.01). Plasma levels of Fg and D-D levels were negatively correlated with PaO2 (r = -0.493, r = -0.438, P < 0.01) before treatment, and also negatively correlated with PaO2 (r = -0.452, r = -0.325, P < 0.01, P < 0.05) after treatment, but they were not significantly correlated with PaCO2 before and after treatment (P >0.05).

CONCLUSIONSQBBAM could play a therapeutic role in improving prethrombotic states of AECOPD patients with BSS. Plasma levels of Fg and D-D were related to the severity of AECOPD.

Acute Disease ; Drugs, Chinese Herbal ; therapeutic use ; Fibrin Fibrinogen Degradation Products ; Fibrinogen ; Hemostatics ; Humans ; Plasma ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; Qi

OBJECTIVETo supply basis for the establishment of quality standard of Cryptolepis buchanaii.

METHODCharacters of crude drugs, microscopic characteristic as well as UV spectrum of the herb were studied.

RESULTLaticifers were found in the cortex and pith of the stem; much papillary non-glandular hair was found covering the stomata in the sub-cuticle of the leaf.

CONCLUSIONThe results can be employed as the basis for identifying the herb.

Cryptolepis ; anatomy & histology ; cytology ; Drug Contamination ; Pharmacognosy ; Plant Leaves ; anatomy & histology ; cytology ; Plant Stems ; anatomy & histology ; cytology ; Plants, Medicinal ; anatomy & histology ; cytology ; Spectrophotometry, Ultraviolet

In-vitro assay methods were established to evaluate transactivation and binding activity of compounds on peroxisome proliferator-activated receptor y (PPARγ). Firstly, plasmids were constructed for transactivation assay of PPARγ response element (PPRE) triggered reporter gene expression, and for cell-based binding activity assay of the chimeric receptor, which was fused with PPARγ ligand binding domain (LBD) and yeast transcriptional activator Gal4. Secondly, by using PPARy competitive binding assay based on time resolved-fluorescence resonance energy transfer (TR-FRET), affinities of compounds and drugs to PPARγ were evaluated. In application of these above methods, the PPARγ activating potency and characteristics of different compounds were evaluated, and a novel benzeneselfonamide derivative, ZLJ01, was found to have comparable binding activity and affinity with the well-known PPARy agonist, but lack of PPRE mediated transactivation activity. In preliminary study on in-vitro hypoglycemic activity, ZLJ1 was found to promote insulin-stimulated glucose uptake by liver cells. Therefore, we believe that combining transactivation and binding activity as well as affinity evaluation, the system could be used to screen non-agonist PPARγ ligand as anovel PPARγ modulator
Genes, Reporter ; Hepatocytes ; Hypoglycemic Agents ; chemistry ; Ligands ; PPAR gamma ; agonists ; chemistry ; Plasmids ; Response Elements ; Sulfonamides ; chemistry ; Transcriptional Activation

Human xanthine oxidase is considered to be a target for therapy of hyperuricemia. Cichorium intybus is a Chinese plant medicine which widely used in Xinjiang against various diseases. In order to screen the inhibitors of xanthine oxidase from C. intybus and to explore main pharmacological actions of cichory a compound collection of C. intybus was built via consulting related references about chemical research on cichory. The three-dimensional crystal structure of xanthine oxidase (PDB code: 1N5X) from Protein Data Bank was downloaded.. Autodock 4.2 was employed to screen the inhibitors of xanthine oxidase from cichory 70 compounds were found to possess quite low binding free energy comparing with TEI (febuxostat). C. intybus contains constituents possessing potential inhibitive activity against xanthine oxidase. It can explain the main pharmacological actions of cichory which can significantly lower the level of serum uric acid.
Chicory ; chemistry ; Databases, Protein ; Drugs, Chinese Herbal ; chemistry ; Enzyme Inhibitors ; chemistry ; Humans ; Molecular Docking Simulation ; Molecular Structure ; Xanthine Oxidase ; antagonists & inhibitors ; metabolism

OBJECTIVETo observe the protective effect of active fractions of Huanglian Jiedu Decoction (HJD) on primary cortical neuron injury after oxygen-glucose deprivation (OGD)/reperfusion (R) injury. Methods Using macroporous resin method, HJDFE30, HJDFE50, HJDFE75, and HJDFE95 with 30%, 50%, 75%, and 95% alcohol were respectively prepared. Then the content of active components in different HJD fractions was determined with reverse phase high-performance liquid chromatography (RP-HPLC). The OGD/R injury model was induced by sodium dithionite on primary cortical neurons in neonate rats. MTT assay was used to observe the effect of four fractions (HJDFE30, HJDFE50, HJDFE75, and HJDFE95) and seven index components of HJD on the neuron viability.

RESULTSRP-HPLC showed active component(s) contained in HJDFE30 was geniposide; baicalin, palmatine, berberine, and wogonside contained in HJDFE50; baicalin, berberine, baicalein, and wogonin contained in HJDFE75. The neuron viability was decreased after OGD for 20 min and reperfusion for 1 h, (P <0. 01), and significantly increased after administered with HJD, HJDFE30, HJDFE50, and HJDFE75 (P <0. 05, P <0. 01). Geniposide, baicalin, baicalein, palmatine, wogonside, and wogonin could increase the cortical neuron viability (P <0. 05, P <0. 01).

CONCLUSIONSHJDFE30, HJDFE50, and HJDFE75, as active fractions of HJD, had protective effect on primary cortical neuron injury after OGD/R. Furthermore, geniposide, baicalin, and baicalein were main active components of HJD.

Animals ; Berberine ; Berberine Alkaloids ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Flavanones ; Flavonoids ; Glucose ; metabolism ; Iridoids ; Models, Animal ; Neurons ; Oxygen ; metabolism ; Rats ; Reperfusion Injury ; drug therapy

A three-dimensional (3D) graphic model of a single-chain Fv (scFv) which was derived from an anti-human placental acidic isoferritin (PAF) monoclonal antibody (Mab) was constructed by a homologous protein-predicting computer algorithm on Silicon graphic computer station.The structure, surface static electricity and hydrophobicity of scFv were investigated. Computer graphic modelling indicated that all regions of scFv including the linker, variable regions of the heavy (VH) and light (VL) chains were suitable. The VH region and the VL region were involved in composing the "hydrophobic pocket". The linker was drifted away VH and VL regions. The complementarity determining regions (CDRs) of VH and VL regions surrounded the "hydrophobic pocket". This study provides a theory basis for improving antibody affinity, investigating antibody structure and analyzing the functions of VH and VL regions in antibody activity.

A three-dimensional (3D) graphic model of a single-chain Fv (scFv) which was derived from an anti-human placental acidic isoferritin (PAF) monoclonal antibody (Mab) was constructed by a homologous protein-predicting computer algorithm on Silicon graphic computer station.The structure, surface static electricity and hydrophobicity of scFv were investigated. Computer graphic modelling indicated that all regions of scFv including the linker, variable regions of the heavy (VH) and light (VL) chains were suitable. The VH region and the VL region were involved in composing the "hydrophobic pocket". The linker was drifted away VH and VL regions. The complementarity determining regions (CDRs) of VH and VL regions surrounded the "hydrophobic pocket". This study provides a theory basis for improving antibody affinity, investigating antibody structure and analyzing the functions of VH and VL regions in antibody activity.

Objective To explore the differences of 2 slice techniques (paste-up method and section method) in the neurosphere slide preparation and the inside structure of the neurosphere. Methods Neural stem cells were isolated from the neonatal mouse telencephalon and the neurospheres on the 7~(th) d of primary culture or subculture were collected. Pasting the intact neurosphere to the slide was the paste-up method. Embedding the spheres with OCT followed by cutting into sections with a frozen section machine was the section method. Immunofluorescence was employed to observe the differences of nestin so as to illustrate the differences of the 2 neurosphere slide preparation methods. Inside structures of the neuroshperes were further studied by HE staining. Results Neural stem cells were successfully isolated and neurospheres formed during in vitro cultures. The superficial cells of the spheres were well stained, but the internal cells could not be stained when paste-up method was adopt; furthermore, the morphology of the cells could not be shown clearly. Both the superficial and internal cells could be well stained with clear morphology when the section method was chosed; as indicated by HE staining, the sphere cells connected to each other and formed complicated cell nets. Conclusion Section method is superior to paste-up method in morphology study of neurosphere and neurosphere enjoys a complicated three-dimension cell growth style.

OBJECTIVETo study surgical techniques for degenerative lumbar scoliosis associated with lumbar stenosis and evaluate their clinical significane.

METHODSThirty-two patients with degenerative lumbar scoliosis associated with spinal stenosis were treated by techniques of posterior lumbar interbody fusion or posterolateral fusion and pedicle screws. There were 18 male and 14 female with 56.8 years old on the average (ranging from 49 to 75 years). There were no evident change of lumberlordosis in 15 cases, and lumber lordosis were obvious loss associated with lumbar subluxation in 17 cases. The correcting, the improvement of back and leg pain, complications and followed-up results were analyzed retrospectively.

RESULTSThirty-two cases were followed-up for 6 to 39 months (the average time of 13 months). The average correction rate of scoliosis was 58.0% and the rate of pain relief was (80.2 +/- 5.8)%. There were two cases of dura sac laceration, two cases of nerve roots injury and a case of pseudoarthritis. During followed-up, correction rate and height of disc spaces were not lost. Shift of interbody cages were no displaced; all the internal fixation got well fusion and the rate of fusion for the bone graft was 96.9%.

CONCLUSIONPosterior pedicle screws combined with interbody fusion or posterolateral fusion is a safe and effective surgical treatment for degenerative lumbar scoliosis associated with lumbar stenosis.

Aged ; Bone Screws ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Scoliosis ; surgery ; Spinal Fusion ; Spinal Stenosis ; complications ; surgery ; Treatment Outcome