Objective To investigate how vascular endothelial growth factor(VEGF) up-regulates intercellular adhesion molecule-1 via protein kinase C(PKC)/ nitric oxide(NO) pathway in the retina of diabetic rats.Methods All the rats were divided into 4 groups: normal,diabetes,diabetes+PKCI and control groups.Diabetes was induced by an intraperitoneal injection of STZ.PKC inhibitor GF109203X was injected intravitreally after 5 months of streptozotocin induced diabetes.NO was determined by nitrate reductase method.VEGF and intercellular adhesion molecule-1(ICAM-1) were measured by Western blot.Results VEGF and NO expressions increased obviously in the retina of diabetic rats compared with those in the normal group(P

Objective To explore the value of time-invariant CTA in assessing collateral circulation of patients with acute ischemic stroke and assisting clinicians in predicting clinical outcomes.Methods The score of collateral circulation was compared between single-phase and time-invariant CTA.NIHSS score was calculated at admission and two weeks after admission.A 50% or greater decrease in NIHSS score over two weeks was considered as major neurologic improvement,which showed good clinical outcome;otherwise,it indicated bad outcome.The predictive ability of time-invariant CTA for clinical outcomes was assessed based on ROC curves.Results Compared with single-phase CTA,more collateral vessels could be viewed on time-invariant CTA.The average score of collateral circulation on time-invariant and single-phase CTA was 1.50±0.69 and 1.15±0.49 respectively (P=0.006<0.05 ).Time-invariant CTA had the moderate predictive ability for clinical outcomes in patients with acute ischemic stroke (AUC=0.810;P=0.032<0.05). Conclusion The time-invariant CTA showed potential value in assessing collateral circulation of patients with acute ischemic stroke and assisting clinicians in predicting clinical outcomes.

Objective To explore the in vitro isolation of ?2m-/Thy-1+ bone marrow-derived liver stem cells(BDLSCs) which bear double features of stem and liver cells from bone marrow stem cells(BMSCs)as so to provide suitable donor cells for the treatment of liver diseases by cellular transplant. Methods ?2m-/Thy-1+ BDLSCs were isolated by magnetic bead cell sorting(MACS) method from cholestatic rats in vitro,and cell purity was detected using flow cytometry.Liver associated phenotype markers were characterized by RT-PCR and immunofluorescence staining. Results BDLSCs isolated by MACS were purified and viable,and possessed hepatocyte-like features at gene and protein levels. Conclusion ?2m-/Thy-1+ BDLSCs are special subsets of BMSCs which may have promising potentials in the stem cell-based treatment of liver diseases.

The study is to develop a method to determine 3 batches leaves of Nauclea officinalis and stems of N. officinalis by HPLC. The differences between strictosamide contents and fingerprints was compared, then chromatographic peak of fingerprints was validated with the assistance of LC-MS. The strictosamide contents in stems of N. officinalis were higher than leaves of N. officinalis. The main chemical composition in leaves of N. officinalis and stems of N. officinalis were alkaloid which revealed by LC-MS. There are 7 chemical compositions were same between them, but the chemical composition in leaves of N. officinalis is more than stems of N. officinalis. This provides a scientific basis for the development of the potential medicinal value of leaves of N. officinalis and the sustainable utilization of N. officinalis.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Rubiaceae ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Nuclear magnetic resonance (1H-NMR) fingerprint of Rhodiola rosea medicinal materials was established, and used to distinguish the quality of raw materials from different sources. Pulse sequence for water peak inhibition was employed to acquire 1H-NMR spectra with the temperature at 298 K and spectrometer frequency of 400.13 MHz. Through subsection integral method, the obtained NMR data was subjected to similarity analysis and principal component analysis (PCA). 10 batches raw materials of Rhodiola rosea from different origins were successfully distinguished by PCA. The statistical results indicated that rhodiola glucoside, butyl alcohol, maleic acid and alanine were the main differential ingredients. This method provides an auxiliary method of Chinese quality approach to evaluate the quality of Rhodiola crenulata without using natural reference substances.
Magnetic Resonance Spectroscopy ; methods ; Principal Component Analysis ; Rhizome ; chemistry ; Rhodiola ; chemistry

After 28 foreign species of AM fungi were inoculated in sterilized soil, the effects of the AM mycorrhizal colonization and the medicine quality of Paris polyphylla var. yunnanensis were observed by combination of inoculation test in pot at room temperature and instrumental analysis. The results showed that, compared with control group (CK), the inoculation of foreign AM fungi in the soil influenced the spore density, mycorrhizal infection rate, and colonization intensity of AM fungi in root system of P. polyphylla var. yunnanensis. The inoculation of foreign AM fungi enhanced the mycorrhiza viability of P. polyphylla var. yunnanensis by increasing the activity of succinic dehydrogenase (SDH) and alkaline phosphatase (ALP) in intraradical hyphae. The content of single steroid saponin in rhizome of P. polyphylla var. yunnanensis showed variation after P. polyphylla var. yunnanensis was inoculated by different foreign species of AM fungi, which was beneficial for increasing the medicine quality; however, the kinds of steroid saponin showed no difference. In a degree, there was a selectivity of symbiosis between P. polyphylla var. yunnanensis and foreign AM fungi. And we found that the Claroideoglomus claroideum and Racocetra coralloidea were best foreign AM fungi species for cultivating P. polyphylla var. yunnanensis under field condition.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Fungi ; classification ; growth & development ; Liliaceae ; chemistry ; growth & development ; microbiology ; Mycorrhizae ; classification ; growth & development ; Plant Roots ; chemistry ; growth & development ; microbiology ; Quality Control

OBJECTIVETo observe the anti-inflammatory effect of total saponins of Panax japonicus on LPS-induced RAW264. 7 macrophages.

METHODThe effect of total saponins of P. japonicus of different concentrations on RAW264. 7 cell viability was determined with the MTT method. The NO kit assay was adopted to detect the NO release of total saponins of P. japonicus to LPS-induced RAW264. 7 cells. The enzyme linked immunosorbent assay (ELISA) was used to detect the secretion of tumor necrosis factor-alpha (TNF-alpha) and interleukin 1-beta (IL-1beta). The reverse transeriptase-polymerase chain reaction (RT-PCR) was used to determine the expression of inducible nitric oxide synthase (iNOS) ,TNF-alpha,IL-1beta. The protein expression of nuclear transcription factor-kappaB p65 (NF-kappaB p65) was tested by Western blot.

RESULTThe safe medication range of total saponins of P. japonicus was less than 80 mg x L(-1). Compared with the LPS model group, total saponins of P. japonicus high, middle and low dose groups (0.1, 1, 10, 40 mg x L(-1)) could significantly reduce the secretion of NO, TNF-alpha, IL-1beta of LPS-induced RAW264. 7 cells, and inhibit the expressions of iNOS, TNF-alpha and IL-1beta mRNA and the protein expression of NF-kappaB p65.

CONCLUSIONThis study preliminarily proves the protective effect of total saponins of P. japonicus on LPS-induced RAW264.7 macrophages. Its action mechanism may be related to NF-kappaB signal pathway.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Inflammation ; drug therapy ; genetics ; immunology ; Interleukin-1beta ; genetics ; immunology ; Lipopolysaccharides ; adverse effects ; Macrophages ; drug effects ; immunology ; Mice ; NF-kappa B ; genetics ; immunology ; Nitric Oxide ; immunology ; Nitric Oxide Synthase Type II ; genetics ; immunology ; Panax ; chemistry ; Protective Agents ; pharmacology ; Saponins ; pharmacology

Silver nanoparticles were synthesized from the extract of Fagopyri Dibotryis Rhizoma and the optimization of synthesis was studied. The absorbance of UV-visible spectroscopy was determined under the different influencing factors such as extracting time of Fagopyri Dibotryis Rhizoma powder, reation temperature of synthesis, volume of Fagopyri Dibotryis Rhizoma extract and concentration of AgNO3 to seek the optimization conditions. By means of FT-IR, TEM, DLS and XRD, the silver nanoparticles were characterized. The results showed that when the boiling time of Fagopyri Dibotryis Rhizoma powder was 5 min, resultant temperature was 25 degrees C, the volume ratio of 0.1 g x mL(-1) Fagopyri Dibotryis Rhizoma extract and 1 mmol x L(-1) AgNO3 was 1 to 10, and the reaction time was 3.5 h, the obtained silver nanoparticles had mean size about 27 nm and Zeta potential about -34.3 mV with good uniformity and dispersivity. Therefore, the green synthesis method of silver nanoparticles using extract of traditional Chinese medicine is stable and feasible.
Fagopyrum ; chemistry ; Light ; Metal Nanoparticles ; chemistry ; ultrastructure ; Microscopy, Electron, Transmission ; Particle Size ; Plant Extracts ; chemistry ; Rhizome ; chemistry ; Scattering, Radiation ; Silver ; chemistry ; Silver Nitrate ; chemistry ; Spectroscopy, Fourier Transform Infrared ; Temperature ; X-Ray Diffraction

BACKGROUND:Previous studies have found that estrogen deficiency causes a reduction in the activity of bone marrow mesenchymal stem cel s (BMSCs). OBJECTIVE:To explore the effect of endothelial progenitor cel s (EPCs) on the BMSCs proliferation and apoptosis ability of osteoporosis rats. METHODS:Healthy female Sprague-Dawley rats, 6 weeks old, were enrol ed and subjected to bilateral ovariectomy to make osteoporosis models. BMSCs and EPCs were isolated using density gradient centrifugation combined with adhesion method, and identified with surface markers, cel proliferation and immunocytochemistry in vitro. We used Transwel inserts to establish EPCs and OVX-BMSCs indirect co-culture system. Control groups were OVX-BMSCs group and sham-BMSCs group in which rats were only subjected to remove the equal amount of fat tissues around the ovary. Flow cytometry was applied to detect BMSCs proliferation and apoptosis ability. RESULTS AND CONCLUSION:Compared with the control groups, the results of flow cytometry test showed that the proportion of OVX-BMSCs at S phase was significantly increased at 3 days of indirect co-culture with EPCs and the apoptosis rate was significanty reduced at 10 days of indirect co-culture with EPCs (both P<0.05). These results suggest that EPCs can promote the proliferation but inhibit the apoptosis of OVX-BMSCs.

BACKGROUND: Current studies on CD62P have focused mainly on cardiovascular diseases, while only few studies have evaluated the effects of CD62P on the development of sepsis and the association between endothelial cell injury with inflammation and coagulation. This study attended to explore the association between endothelial cell injury with inflammation and coagulation by evaluating the expression of soluble CD62P (s-CD62P) in plasma and its mechanism in patients with sepsis, thus to provide the evidence of effective treatment of sepsis with anti-adhesion therapy targeted CD62P. METHODS: A total of 70 critically ill patients with systemic inflammatory response syndrome (SIRS) admitted to intensive care unit (ICU) between September 2009 and February 2010 were enrol ed for a prospective and control study. According to the diagnostic criteria of sepsis/SIRS, the patients were divided into two groups: a sepsis group (n=38) and a SIRS group (n=32). Another 20 healthy volunteers served as a control group. Patients in the sepsis group and SIRS group were matched by clinical signs of high blood pressure, diabetes and its complications. The demographics of the patients including age, sex, body mass index (BMI), smoking and alcohol addict were compared among the groups. Six mL peripheral blood samples were collected within 24-hour admission in ICU for enzyme-linked immunosorbent assay (ELISA) to detect the plasma levels of s-CD62P, TNF-α, and hs-CRP. And variables of coagulation function such as platelet (PLT), prothrombin (PT), activated partial thromboplastin time (APTT), D-dimer and antithrombin-III (AT-III) were analyzed during 24 hours after admission to ICU. Meanwhile sequential organ failure assessment (SOFA) score of critically ill patients was evaluated. Data were expressed as mean±standard deviation and were statistical y analyzed by using SPSS 17.0 statistical software. The differences in plasma levels of s-CD62P of patients in each group were analyzed by ANOVA and the Kruskal-Wallis test. The relations between s-CD62P and inflammatory cytokines as well as with coagulation were determined by Pearson's product moment correlation coefficient analysis. Changes were considered as statistically significant if P value was less than 0.05. RESULTS: Compared with the control group and SIRS group, the sepsis group demonstrated significantly higher levels of s-CD62P, TNF-α and highly sensitive C-reactive protein (hs-CRP) (P<0.05). The plasma levels of D-dimer, PT, and APTT in the sepsis and SIRS groups were significantly higher than those in the control group, while the platelet count and the activity of AT-III were obviously lower (P<0.05). In the sepsis group, the plasma levels of hs-CRP and TNF-α were positively correlated with PT, APTT, and D-dimer, and negatively correlated with AT-III and PLT (P<0.05). The plasma levels of s-CD62P were significantly correlated with the plasma levels of TNF-α, hs-CRP, D-dimer, PT, and APTT, whereas they were correlated negatively well with PLT and AT-III (P<0.05). CONCLUSIONS: The concentration of plasma s-CD62P is elevated as a early biomarker in patients with sepsis, and it serves as one of the pathogenic factors responsible for endothelial cell damage. Coagulation and mediators of inflammation promote each other, aggravating the severity of sepsis. Plasma s-CD62P may be an important factor for the development of coagulation and inflammatory reaction.