Objective To isolate and identify the adult neural stem cells from the parenchyma of spinal cord in adult mouse.Methods The parenchymal spinal cord from adult mouse was dissected and dissociated by mechanical trituration.The tissue suspension was cultured in serum-free DMEM/F12 medium supplemented with EGF and B27.The cell colonies generated from a single cell were screened by limited dilution and incubated with BrdU.The cell colonies were transferred into medium with serum to induce differentiation.The cells were identified with antibodies to Nestin,BrdU,MAP2 and GFAP by immunofluorescence staining.Results The cells were cultured for seven days to generate proliferative neurospheres.The majority of cells in these neurospheres expressed Nestin and were differentiated into MAP2-positive cells and GFAP-positive cells in medium containing with fetal bovine serum.Conclusion A significant number of neural stem cells are present in the parenchymal adult mouse spinal cord and can proliferate and also give rise to neurons and glia in vitro.

Obesity is an established risk factor for endometrial cancer. Leptin, a secreted protein of the ob gene by white adipose tissue, plays an important role in the regulation of food intake and energy consumption in the brain and acts as a potential growth stimulator in normal and neoplastic cancer cells. However, a direct role for leptin in endometrial cancer has not been demonstrated. In the present study, the effect of leptin on the proliferation of Ishikawa endometrial cancer cells was investigated as well as the possible mechanism(s) underlying this action in endometrial cancers which express both short and long isoforms of leptin receptors. The expression of leptin receptor (ObRb) in Ishikawa cells was detected by RT-PCR and Western blotting. The cells after serum starvation, were treated by leptin with various concentrations (0, 10, 50, 100, 150 ng/mL) for different durations (6, 12, 24 h). The effect of leptin treatment on cell proliferation was examined by MTT assay. Meanwhile, inhibitory effect of Janus tyrosine kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) inhibitor AG490 or extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor PD98059 on the proliferation of Ishikawa cells induced by leptin was also studied. Ishikawa cells were treated with 100 ng/mL leptin for various periods (0, 20, 40, 60 min), and the levels of STAT3 phosphorylation and ERK1/2 phosphorylation were examined by Western blotting. The results showed that leptin induced the phosphorylation of STAT3 and the activation of ERK1/2 in a time- and dose-dependent manner in the Ishikawa endometrial cancer cells. Blocking STAT3 phosphorylation with the inhibitor AG490, or blocking ERK1/2 activation by the specific ERK1/2 kinase inhibitor, PD98059, abolished leptin-induced proliferation of Ishikawa cells. In addition, leptin was found to potently induce the invasion of endometrial cancer cells in a Matrigel invasion assay. Leptin-stimulated invasion was effectively blocked by pharmacological inhibitors of STAT3 (AG490) and ERK1/2 kinase (PD98059). These results suggested that leptin promotes endometrial cancer growth and invasiveness by activating STAT3 and ERK1/2 signaling pathways and therefore blocking its action at the receptor level can be a rational therapeutic strategy.

Objective To determine the serum level of chernokine CCL27 in patients with psoriasis vulgaris,and to analyse its clinical relevance.Methods A total of 61 patients(40 in progressive stage and 21 in stable stage)with psoriasis vulgaris,with an average disease duration of 37.97±14.34 years,were included in this study.Appropriate thempy was given to these patients.Serum samples were collected from the patients before and after therapy,as well as from 45 healthy human controls.ELISA was applied to examine the serum concentration of CCL27.Clinical severity of psoriasis vulgaris was assessed by psoriasis area and severity index(PASI)score.Results Serum level of CCL27 was 670.02±262.15 ng/L in psoriatic patients,compared to 373.10±92.84 ng/L in the controls(t=8.18.P＜0.01).Increased serum level of CCL27 was observed in patients with progressive psoriasis vulgaris compared to those with stable psoriasis (799.94±214.54 ng/L vs 422.57±135.53 ng/L,t=8.39,P＜0.01).After 8 weeks of therapy,a significant decrease was noticed in the serum level of CCL27 in patients who experienced≥70%reduction in PASI score(t=9.95,P＜0.01).but not in those experiencing a PASI reduction of＜70%(t=1.84,P＞0.05).The serum level of CCL27 was positively correlated with PASI score(r=0.58,P＜0.01).Conclusions The serum level of CCL27 is significantly elevated in patients with psoriasis vulgaris,and it is correlated with the disease severity.

One new neolignan identified as 2, 3-( trans) -dihydro-2-(4-hydroxy-3-methoxyphenyl) -3-[(beta-D-glucopyranosyloxy) methyl]-7-methoxybenzofuran-5-propenoic acid (1) and five known steroidal glycosides namely torvoside A(2), torvoside C(3), torvoside H(4), solanolactoside A (5), (25S)-6alpha-hydroxy-5alpha-spirostan-3-one-6-0-[alpha-L-rhamnopyranosyl-(1-->3-beta3)-beta-D-D-quinovopyr-anoside] (6) were isolated from the fruits of Solanum torvum. Their structures were elucidated on the basis of 1D, 2D NMR and MS spectroscopic analysis.
Fruit ; chemistry ; Isomerism ; Lignans ; chemistry ; isolation & purification ; Solanum ; chemistry

Objective To observe myocardial tolerance to ischemia/reperfusion (I/R) injury in rats after exposure to X-ray irradiation.Methods Twelve male rats were randomly divided into control group and radiation group.The rat model of radiation-induced heart disease was established in the radiation group by precordial irradiation with 20.0 Gy of 6 MV X-ray in a single fraction.At 14 days after model establishment,the Langendorff perfusion technique was performed in the two groups and the cardiac parameters including left ventricular developing pressure (LVDP),left ventricular end diastolic pressure (LVEDP),maximal rate of left ventricular pressure rise/fall (+/-LVdp/dtmax),and coronary flow (CF)were recorded.Myocardial infarct size after I/R was compared between the two groups by 2,3,5-triphenyltetrazolium chloride staining.Results After 30 minutes of ischemia and 60 minutes of reperfusion,the irradiation group had a significantly slower CF than the control group (5.64±0.35 vs.8.38±0.52 ml/min,P=0.002).Moreover,the irradiation group had substantially poorer recovery of cardiac function in isolated hearts compared with the control group,as shown by a significantly reduced LVDP (25.4±2.31 vs.52.76±2.76 mm Hg(1 mm Hg=0.133 kPa),P=0.000),significantly reduced+/-LVdp/dtmax(547.04±78.74 vs.1 100.05±83.35 mm Hg(1 mm Hg=0.133 kPa)/s,P=0.001;-408.81±56.74 vs-813.62±73.82mm Hg(1 mm Hg =0.133 kPa)/s,P=0.002),and a significantly increased LVEDP (85.29±4.61 vs.65.65±3.65 mm Hg (1 mm Hg =0.133 kPa),P=0.012).X-ray irradiation induced a significantly increased percentage of myocardial infarct size in rats (44.67％±0.95％ vs.30.46％±0.96％,P=0.000).Conclusions X-ray irradiation can induce coronary injury,reduce myocardial tolerance to I/R injury,and increase myocardial infarct size after I/R in rats.

ObjectiveTo observe the injured changes of brain myelin sheath structure and myelin basic protein (MBP) content induced by amyloid β peptide (Aβ) and effect of GETO on these changes.MethodsThe experimental rat model of Alzheimer's disease was established with Aβ1-42 injection into hippocampus. 4 weeks later, the myelin sheath structure of the CA1 area of the rat hippocampus was taken and observed by electromicroscope, and distribution and content of MBP were examined with immunohistochemical method.ResultsThe electromicroscope showed that the structure of myelin sheath became relaxing, disorder, homogenization and default of hippocampus CA1 in the model rats. In GETO treated group, the structure of myelin sheath was integrity and continuum. Immunohistochemical test showed that the staining and numbers of myelin sheath of model rats was thinner than that of normal rats and GETO treated rats. The numbers, mean area and mean density of positive staining axon in hippocampus CA1 of MBP in the model rats were significantly different from those in the normal group and GETO group (P<0.01).ConclusionAβ1-42 injection into hippocampus in rats can impair myelin sheath to make MBP release and GETO can ameliorate these changes.

BACKGROUNDIn steroid-induced osteonecrosis, hypertrophy and hyperplasia of marrow fat cells and lipid deposition of osteocytes can be found in the femoral head. However, the precise reason is not clear yet. The aim of this study was to observe the effect of dexamethasone (Dex) on differentiation of marrow stromal cells (MSCs), and to investigate the pathobiological mechanism of steroid-induced osteonecrosis.

METHODSMSCs in cultures were treated with increasing concentrations of Dex (0, 10(-9), 10(-8), 10(-7), and 10(-6) mol/L) continuously for 21 days. The cells, which were exposed to 0 mol/L (control) or 10(-7) mol/L Dex for 4 - 21 days, were then cultured for 21 days without Dex. MSCs were stained with Sudan III. Number of adipocytes was counted under a light microscope. The activity of alkaline phosphatase (ALP) of MSCs treated with 0, 10(-8), 10(-7), and 10(-6) mol/L Dex for 12 days, and that treated with 0 mol/L and 10(-7) mol/L Dex for 8, 10, or 12 days were determined. The levels of triglycerides, osteocalcin and cell proliferation of MSCs treated with 0 mol/L and 10(-7) mol/L Dex were detected. The mRNA expression levels of adipose-specific 422 (aP2) gene and osteogenic gene type I collagen in MSCs treated with 0 mol/L and 10(-7) mol/L Dex for 6 days were analyzed by whole-cell dot-blot hybridization. Statistical analysis was performed using Student's t test and analysis of variance. P values less than 0.05 were considered significant statistically.

RESULTSThe number of adipocytes in cultures increased with the duration of MSCs' exposure to Dex and the concentration of Dex. The level of ALP activity in the MSCs decreased with concentration of Dex. In the control group, it was 8.69 times of that in the 10(-7) mol/L Dex group on day 12 (t = 20.51, P < 0.001). The level of triglycerides in 10(-7) mol/L Dex group was 3.40 times of that in the control (t = 11.00, P < 0.001). The levels of cell proliferation and osteocalcin in the control were 1.54 and 2.42 times of that in the 10(-7) mol/L Dex group respectively. As compared to the control, the mRNA expression of adipose-specific 422 (aP2) gene in 10(-7) mol/L Dex group was significantly increased (t = 36.48, P < 0.001), and that of osteogenic gene type I collagen was decreased (t = 42.07, P < 0.001).

CONCLUSIONSDex can directly induce the differentiation of MSCs into a large number of adipocytes and inhibit their osteogenic differentiation, which provide a novel explanation for the pathologic changes of steroid-induced osteonecrosis.

Adipogenesis ; drug effects ; Alkaline Phosphatase ; metabolism ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dexamethasone ; toxicity ; Female ; Mice ; Osteocalcin ; genetics ; Osteonecrosis ; chemically induced ; RNA, Messenger ; analysis ; Radioimmunoassay ; Stromal Cells ; cytology

Adolescent ; Brain Diseases ; diagnostic imaging ; Brain Neoplasms ; diagnostic imaging ; surgery ; Diagnosis, Differential ; Hematoma, Epidural, Cranial ; diagnostic imaging ; Hernia ; diagnostic imaging ; Humans ; Male ; Plasmacytoma ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed

OBJECTIVETo observe the intervention effect of acupuncture on early onset of selec- tive serotonin reuptake inhibitors (SSRIs) in treating depressive disorder, and to study its effect on ser- um 5-HT and unbalanced inflammatory cytokines secreted by TH1/TH2.

METHODSTotally 90 patients with depressive disorder were randomly assigned to the drug control group (as the control group, 45 cases) and the acupuncture combined drug treatment group (as the treatment group, 45 cases). All patients were treated for 4 consecutive weeks. Another 45 healthy subjects were recruited as a healthy control group. The effect of acupuncture on early onset of SSRls in treating acute phase depressive disorder pa- tients was evaluated by HAMD score in the control group and the treatment group before treatment,and at weekends of the 1st, 2nd, and 4th week after treatment. Besides, their serum levels of 5-HT, IL-1β and IL-6 (secreted by TH1), and IL-4 and IL-10 (secreted by TH2) were detected before treatment and after treatment at the weekend of the 4th week.

RESULTSCompared with the healthy control group,serum lev- els of 5-HT, IL-4, and IL-10 decreased in the two drug-treated groups before treatment (P < 0.01); serum levels of IL-1β and IL-6 increased (P <0.01). Compared with before treatment in the same group, HAMD score decreased in the control group at weekends of the 2nd and the 4th week after treatment (P < 0.01); HAMD scores decreased in the treatment group at weekends of the 1st, 2nd, 3rd,and 4th week after treatment (P < 0.01); serum levels of 5-HT, IL-4, and IL-10 increased,serum levels of IL-1β and IL- 6 decreased in the two drug-treated groups after treatment (all P < 0.01). Compared with the control group at the same time point,HAMD scores decreased in the treatment group at weekends of the 1st, 2nd,3rd,and 4th week after treatment (P < 0.01),serum levels of 5-HT, IL-4, and IL-10 increased (P < 0.05, P < 0.01), serum levels of IL-6 decreased (P < 0. 01).

CONCLUSIONAcupuncture could accelerate early onset of SSRIs in treating acute phase depressive disorder, and effectively regulate serum 5-HT levels and inflammatory cytokines secreted by TH1/TH2.

Acupuncture Therapy ; Cytokines ; Depressive Disorder ; drug therapy ; Drugs, Chinese Herbal ; Humans ; Interleukin-10 ; Interleukin-1beta ; Interleukin-4 ; Interleukin-6 ; Serotonin Uptake Inhibitors ; therapeutic use

Objective To investigate the characteristics of renal hemodynamic and the esoteric prostacyclin(PGI2),thromboxane A2(TXA2)level in children with early Henoch-Schonlein purpura(HSP),and study the function of TXB_2/6-Keto-prostaglandin F(6-Keto-PGF_(1?))(T/K)numerus in early changes of kidney injury.Methods Children involved in the experiment were dicided into 3 groups.Thirty-one patients with HSP,divided into 2 groups according to routine urianlysis:children with HSP without renal damage group(n=16)and Henoch-Schonlein purpura nephritis(HSPN)group(n=15).Control group with 16 healthy children,their age and sex match with the other 2 groups.The urine of all children,including the children in control group,was sampled in 24 hours.The urinary production of the samples were kept in the freezer at-20 ℃.The radioimmunoassay was applied to determine the 6-Keto-PGF_(1?),TXB_2 quantitatively,and calculate the number of T/K.In the early morning the children accept the Doppler arteria renalis sonography with an empty stomach to determine the Vmax of the period of contraction of the arteria renalis the Vmin of diastolic phase and the resistent index(RI).SPSS 13.0 software was used to analyze the data.Results 1.The renal hemodynamic indicated a change of high velocity and resistance,the masculine rate(83.9%)was ob-viously higher than that in routine urinalysis(48.4%)(?2=5.79 P0.05).The RI in the former group(0.798?0.165)was much higher than that in the other one(0.637?0.116)(t=4.02 P