Objective To investigate the role of ceramide pathway in cell proliferation and early apoptosis induction in U251 glioma cell after cannabinoid receptora agent anandmide(AEA)treatment.Methods U251 gliom cells were treated with AEA(1-10 μmol/L),Ceramide(5-20 μmol/L) and fumonisin B1 (FB1) (10 μmol/L) pretreatment.The growth inhibition rate of U251 was investigated by MTT assay.The early events of the apoptosis were measured by flow cytometry using annexin-V/propium iodide(PI) double staining method.Results Different concentrations of AEA inhibited the proliferation of human glioma U251 cells,and had synergistic effect with CM by FB1(10 μmol/L)pretreatment for 24 h.After exposure to AEA(10 μmol/L)for 24 h,U251 gliomacells could undergo the early cell apoptosis which was affected by FB1(10 μmol/L).Conclusion AEA through the CM de novo synthesis pathway,and CM concentration was lazy in collaboration,thus inhibiting human glioma U251 cell proliferation and induce early apoptosis.

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.

According to the TCM theory, human growth, development and aging are closely related to kidney function and the main cause of human senility is deficiency of “kidney essence”. Recent studies showed that the “kidney” in TOM include the regulatory function of the neuroendocrine and immune systems.
In this paper, the effect of Guzhen Recipe (_??__??__??_), an aging retarding prescription composed of kidney tonifying herbs such as Radix Polygoni Multiflori (_??__??_), Herba Cistanchis (_??__??_) Rubus Chiagii (_??__??__??_) etc. on the hippocampus and HPAT axis in senile rats were observed. The results showed : (1) With age the number of glucocorticoid receptors (GCR) in the hippocampus, the concentrations of NE, DA, 5-HT in the hypothalamus were markedly decreased, while the plasma corticosterone concentration in senile rats was obviously increased. (2) The thymocyte GCR nuclear transition rate was increased significantly ; (3) The contents of cytoplasmic protein, nuclear RNA and DNA, the thymus weight/body weight ratio, the serum thymic factors as well asthe rate of lymphocyte transformation, the activity of IL-2 and NK cells in senile rats were obviously decreased compared to those of young control rats. Guzhen recipe however, was capable of improving these changes in the neuroendocrine-thymic net work in senile rats to varying degrees, suggesting that the kidney with Chinese Medicine might retard the functional degeneration of thymic endocrine partly by improving the feedback regulation of the hippocampal GCR and increasing the concentrations of NE and 5-HT of hypothalamus, thus decreasing the plasma corticosterone ; Simultaneously weakening the nuclear transition rate of thymocyte GCR, which in turn improving the immune function of the senile animal.
We concluded that Guzhen Recipe may retard the functional degeneration of both the hypo-thalamuspituitary-gornadal-thymic and hypothalamus-pituitary-thyroidal-thymic axis. All the above results revealed that retarding the aging effect using TCM in volves multiple-leves, multiple-systems and links. Using holistic regulation, imbalance links in aging process are modulated and the equilibrium of Yin and Yang are maintained. This is an important characteristic of the TCM to retard aging.
This research was supported by funds from National Natural Science Foundation of China No. 38970908.

Objective To analyze the distribution of class 1,2 and 3 integrons and their gene cassettes,and to explore its relationship with drug resistance in Shigella.Methods Antimicrobial susceptibility was detected by minimal inhibitory concentration (MIC) method.All the genes of integrons and gene cassettes were amplified by polymerase chain reaction (PCR).The amplicons were identified by amplified fragment length polymorphism (AFLP) and sequencing.Results Fifty seven multi-drug resistant strains were identified from a total of 62 Shigella strains (91.9％).Among multi drug resistant strains,52 strains carried integrons of class 1 (91.2 ％) and 55 strains carried integrons of class 2 (96.5％).Only 2 strains carried class 1 integrons alone,5 strains carried class 2 integrons alone and 50 strains had both class 1 and class 2 integrons.Class 3 integrons were not detected.The gene cassettes of typical class 1 integrons,dfrV and dfrA17-aadA5,were detected in 6 strains and 2 strains,respectively.Atypical class 1 integrons with gene cassettes blaOxA30 aadA1 were detected in 44 strains.The typical and atypical class 1 integrons coexisted in 6 Shigella flexneri strains.Gene cassettes for class 2 integrons were dfrA1 sat1-aadA1.Conclusions The multi-drug resistant Shigella strains are widely distributed in Ji'nan,and the atypical class 1 integrons and class 2 integrons are common in these strains.Coexistence of the two integrons is observed in some of the strains.

0.05). ConclusionThe present study confirms that tiotropium 18 ug once daily compare with ipratropium can reduce lung hyperinflation,ease exertional dyspnea and improve symptom-limited exercise tolerance in COPD patients.

Objective To investigate the efficacy of computer-assisted postural balance training system combined with Bobath balance training on balance disorders after cerebellar stroke.Methods Forty patients with balance disorders after cerebellar stroke were randomly divided into either a combined training group or a control group (n =20 in each group).The control group was trained with Bobath balance training and routine rehabilitation treatment.On the basis of this,the combined training group used the computerassisted postural balance training system for rehabilitation treatment.The balance function,activities of daily living and motor function were evaluated with the Berg Balance Scale (BBS),modified Barthel Index,mBI)and Fugl-Meyer Assessment (FMA) before and after treatment,and the computer-assisted postural balance training system was used to conduct the postural balance ability according to the center of gravity track length,rectangle area of track,and peripheral area of track.Results There were no significant differences in various indexes before treatment between the patients of both groups.The scores of BBS,mBI and FMA after treatment were increased obviously (all P ＜ 0.001).The the center of gravity track length,rectangle area of track,and peripheral area of track were decreased compared before treatment (P ＜0.001 or 0.05).In addition to the FMA score,all the indexes of the combined training group after treatment were significantly better than those of the control group (all P＜0.05).Conclusions On the basis of Bobath balance training,using computer-assisted postural balance training system can obviously improve the balance disorder and extremities motion ability after cerebellar stroke,and thus improving the activities of daily living in cerebellar stroke patients.

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

Objective Post-transcription RNA interference (RNAi) is more and more widely applied in multigene therapy.This study aimed to establish an subcutaneous xenotransplanted tumor (SXT) model of human HT-29 colon carcinoma in nude mice and investigate the effects of the survivin shRNA-APC double gene co-expression lentiviral vector on the growth of SXT.Methods Thirty-five nude mice were equally divided into five groups, double-gene survivin shRNA, survivin shRNA, APC, empty vector, and blank, and injected into the left anterior axillary with respective stably transfected cell lines and human HT-29 colon carcinoma cells, all at 2×106/mL, to establish an SXT model of human HT-29 colon carcinoma.The inhibition rate of tumor growth was calculated by measuring the size and weight of the SXT, the expressions of survivin mRNA and protein in the tumor tissue detected by real time PCR and immunohistochemistry respectively, and the apoptosis of the HT-29 colon carcinoma cells determined by TUNEL.Results The mean size and weight of the SXT were significantly reduced in the double-gene survivin shRNA-APC, survivin shRNA, and APC groups as compared with the blank and empty vector groups (P<0.05), though increased in the survivin shRNA and APC groups in comparison with the double-gene group (P<0.05).The expressions of survivin mRNA and protein in the tumor tissue were remarkably lower in the double-gene survivin shRNA-APC, survivin shRNA, and APC groups than in the blank and empty vector groups (P<0.05), even lower in the double-gene group than in the survivin shRNA, and APC groups (P<0.05).The apoptosis rate of the HT-29 colon carcinoma cells was markedly up-regulated in the double-gene survivin shRNA-APC ([56.72±3.17]%), survivin shRNA ([33.64±2.03]%), and APC groups ([31.19±1.79]%) as compared with the blank ([9.89±0.31]%) and empty vector groups ([10.06±0.43]%) (P<0.05), even more significantly in the double-gene than in the survivin shRNA and APC groups (P<0.05).Conclusion The survivin shRNA-APC double gene co-expression lentiviral vector can reduce the expression level the survivin gene, promote the apoptosis of colon carcinoma cells, and suppress the growth of the subcutaneous xenotransplanted tumor.

Aim To determine the feasibility of transforming growth factor-a ( TGF-a ) as a diagnostic bio-marker for systemic-to-pulmonary shunts induced pulmonary arterial hypertension ( PAH ). Methods Systemic-to-pulmonary shunts induced PAH was built by combined surgery ( right pulmonary artery was ligated and a cervical shunt was established one week later). A total of 49 patients with congenital heart diseases were recruited, including 24 congenital heart disease patients without PAH(CHD) and 25 congenital heart disease patients with pulmonary arterial hypertension (CIID-PAH). Moreover, 20 healthy volunteers matched by age and sex were also included. Enzyme linked immunosorbent assay (EL1SA) was used to test TGF-a concentrations in plasma of systemic-to-pulmonary shunts induced PAH rats and CHD-PAH patients. Results ELISA results showed that TGF-a levels in plasma of systemic-to-pulmonary shunts induced PAH rats were significantly higher than those of sham operated group. Spearman correlation analysis showed that plasma TGF-a concentrations were positively associated with right ventricular systolic pressure, pulmonary arterial systolic pressure, mean pulmonary arterial pressure and right ventricular hypertrophy index. The plasma concentration of TGF-a in CHD-PAH patients was much higher than that of CHD patients and healthy vol- unteers ( CON); however, there was no significant difference between CHD group and CON group; Using 314 ng • L"1 as cutoff value of TGF-a for the diagnosis of CHD-PAH, the sensitivity, specificity and area under the cure was 0. 760, 0. 750 and 0. 895, respectively. Conclusions Plasma concentration of TGF-a increases with the progression of systemic-to-pulmonary shunt induced PAH; the level of TGF-a in plasma may be a potential biomarker for the diagnosis of systemic- to-pulmonary shunt induced PAII.

OBJECTIVEHigh altitude pulmonary edema (HAPE), a life-threatening disease, has no biological markers used for the routine prevention, diagnosis and treatment. The aim of this study was to identify serum proteins differentially expressed in patients with HAPE for discovering essential biomarkers.

METHODSA complete serum proteomic analysis was performed on 10 HAPE patients and on 10 high altitude and 11 sea level healthy people as control using two-dimensional gel electrophoresis, followed by matrix-assisted laser desorption/ionization mass spectrometry and peptide mass fingerprinting. Finally, two most significantly changed proteins were validated by enzyme-linked immunosorbent assay (ELISA).

RESULTSEight protein spots stained with differential intensity, respresenting 5 distinct proteins were identified in patients compared with healthy controls through analysis of these composite gels. Among them, four proteins, namely alpha 1-antitrypsin(alpha1-AT), Haptoglobin(Hp), apolipoprotein A-1 (apoA-1) and Complement C3 increased remarkably, while one protein, apolipoprotein A-IV (apoA-IV) decreased significantly. The variation of alpha1-AT and Haptoglobin, as detected by ELISA, was consistent with the results from proteomic analysis.

CONCLUSIONSIt is well known that Hp, alpha1-AT and complement C3 are associated with inflammation and apoA-1 and apoA-IV play important roles in lipid absorption, transport and metabolism. Therefore, the significant expression changes of Hp, alpha1-AT and complement C3 and apoA-1 and apoA-IV between HAPE patients and their corresponding healthy controls highlight the role of inflammatory response system and lipid metabolism system in the pathophysiology of HAPE.

Altitude ; Biomarkers ; blood ; Blood Proteins ; metabolism ; Case-Control Studies ; Electrophoresis, Gel, Two-Dimensional ; Enzyme-Linked Immunosorbent Assay ; Humans ; Peptide Mapping ; Proteome ; Pulmonary Edema ; blood ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization