Objective To establish an in vitro oxygen-glucose deprivation/reoxygenation axon injury model in rat hippocampal neurons in order to found a basis for research concerning the injury of oxygen glucose deprivation and axons regeneration. MethodsThe hippocampal neurons isolated from rats 24 h after born and cultured for 7 d were exposed to D-hanks solution with nitrogen gas instead of the original culture medium for 0.5, 1.0 and 1.5 h respectively, and then continue to be cultured in the original culture medium with oxygen. LDH content in the culture media was measured at 1, 5, 24, 48 and 72 h after reoxygenation. The morphological changes of neuron and axon were observed with inverted phase contrast microscopy. ResultsAfter oxygen-glucose deprivation/reoxygenation injury, hippocampal neurons became darker, swollen, and were with shorten axons. With the elapse of time, the LDH content was increased. The survival rate of hippocampal neurons was higher and the change of axon length was more obvious in the group of oxygen-glucose deprivation for 0.5 h than in the other groups. ConclusionAn in vitro oxygen-glucose deprivation/reoxygenation axon injury model in rat hippocampal neurons is successfully established.

Objective To explore the dysfunction of dendritic cells (DC) related to TGFβ reversed after blocking the TGFβ signal pathway by recombinant adenovirus vector encoding for Smad7.Methods Smad7 by recombinant adenovirus vector was transfected into dendritic cells.Expression of immunologic phenotypes was detected by FCM,and CTL activity induced by DC was compared.Results The DC modified with Smad7 still expressed high adhesiveness factor related to maturation even if existing exogenous TGFβ1,which was significant statistically compared with DC transfected with control adenoviral vector (P ＜0.01).Even if existing exogenous TGFβ1,the DC modified with Smad7 pulsed with soluble antigen associated with Lewis pulmonary carcinoma could still induce potent CTL activity against Lewis pulmonary carcinoma,which showed significant difference with DC-Ad-c (P ＜0.01).Conclusion The inhibitory effects on function of DC of TGFβ may be reversed by blocking the Smad signal of TGFβ pathway.

Objective To evaluate surgical therapy for recurrent rectovaginal fistula.Methods In this study.two patients were treated by endorectal advancement flap repair and one patient was treated by vascular pedicled segment of small bowel.Results All patients were cured and followed up from 4 to 20 months.During the period of follow-up there was no recurrence.Conclusions The procedures and timing of operation are important factors for a successful repair.Both the endorectal advancement flap and patch of intestine provide an effective methods in repairing recurrent rectovaginal fistula.

Objective:To explore the effect of repetitive high-frequency magnetic stimulation (H-F rTMS) of the dorsolateral part of the prefrontal cortex (DLPFC) combined with smoking-related cues on nicotine addicts′ cigarette craving, the concentration of exhaled CO and sleep quality.Methods:Sixty nicotine addicts were randomly divided into groups A, B and C, each of 20. All were given H-F rTMS five times a week for two weeks, while those in groups A and B watched smoking and non-smoking pictures for ten minutes, respectively. Before and after the intervention, all of the subjects self-reported their cigarette cravings using a visual analogue scale. Exhaled CO (CO ppm) was measured and the Pittsburgh sleep quality index (PSQI) was evaluated. Results:After the intervention the average craving score, CO ppm and PSQI score had improved significantly in all three groups. The average craving score and CO ppm of group A were both significantly better than in the other two groups. Conclusions:rTMS can significantly improve cigarette craving, CO ppm and sleep quality of cigarette adicts. Viewing smoking-related pictures as an addition to rTMS can even better the effects of rTMS.

Animals ; Antibodies, Bacterial ; blood ; Campylobacter Infections ; complications ; microbiology ; Campylobacter jejuni ; isolation & purification ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Fatal Outcome ; Humans ; Male ; Polyradiculoneuropathy, Chronic Inflammatory Demyelinating ; etiology ; Swine

Objective To investigate the correlation of PICC tip position and weight gaining in very low birth weight infants.Methods We performed a retrospective study using chest X-ray films of very low birth weight infants in NICU who had PICCs inserted in a tertiary hospital.We recorded the tip positions on plain radiographs and calculated the position change,and calculated weight gaining ratio.Spearman regression model was used to analyze the relationship between tip position migration and weight gaining ratio.Results A total of 57 cases of very low birth weight infants were included,containing 246 X-ray films.On the first day of taking X-ray,weight was 0.6-1.46 (1.06±0.25) kg,the median and interquartile spacing are 5(3,7)d.The last time of taking X-ray was(24.0±9.1) dafter PICC placement;weight gaining ratio was 11.8％～114.8％,the median and interquartile spacing are 41.5％ (27.1％,65.3％).All PICCs tip position changed,75％ of which migrated more than 2 vertebrae,50％ of which migrated 3 vertebrae,and the median and interquartile spacing are 3 (1.8,3.5) vertebrae.The distances of tip migration were correlated with weight gaining ratio.With an approximate 2,3 and 4 vertebrae of PICC tip migration,the corresponding weight gaining ratio was about 40％,70％ and 100％.The correlation coefficient between PICC tip position migration and weight gaining ratio was-0.7(P＜0.01),but there was difference in different insertion sites.Conclusion PICC tip position is greatly influenced by weight gaining among very low birth weight infants.By considering the initial placement position,the crucial moment to assess catheter location is at 40％ and 70％ weight gaining ratio.After 100％ weight gaining,PICCs should be removed or replaced.PICCs can easily be affected by bone growth and limb movement,require higher frequency of catheter localization.

Objeetive To study the differences of the biological characteristics and immune regulation function of adipose mesenchymal stem cells (AMSCs)from psoriasis patients and healthy people.Methods AMSCs were isolated and cultured from human psoriatic and healthy adipose tissue,the phenotypes and cell cycle of AMSCs taken from three generation were detected by flow eytometry.Alkaline phosphate enzyme staining and oil red o staining were used respectively to identify their adipogenic and osteogenic capacity.Next,the levels of inflammation antimicrobial proinflammatory factor were detected by PCR and ELISA.Then gene expression profile of AMSCs were screen by gene expression profile chip,as so to bolting the the gene array related with immunology gene.Results There was no significant change in cell morphology,and cell surface markers were expressed high for CD29,CD44,CD73,while lower for CD31,CD45 and HLA-DR.AMSCs of psoriasis patients and healthy people both had the ability of adipogenic and osteogenic differentiation.But the cell cycle showed the third generation AMSCs proliferation rates were slower than that of normal control,as compared with healthy controls,adipogenic differentiation ability was stronger.What'more,the level of inflammatory cytokines in psoriasis group was lower than that in controls such asIL-10,IDO,TGF-β,on the contrary the levels of proinflammatory factor in psoriasis group were higher than that in controls,such as TNF-α,IFN-γ.In addition,gene chip results suggested that psoriasis group AMSCs had obvious expression differences on JAK-STAT pathway with healthy controls.Conclusions Compared with the control,there are significant differences in patients AMSCs proliferation and adipogenic differentiation ability,immune inflammation suppression control ability is weaken,this phenomenon may be associated with JAK-STAT immune pathways related to downgrade.

The chemical structures of P1 A was identified by complete acid hydrolysis, partial acid hydrolysis, periodate oxidation-Smith degradation, methylation analysis, IR and NMR. The results showed that P1 A had a backbone consisting rhamnose, mannose, glucose and galactose. The side chain possessed arabinose and xylose. 1-->, 1-->6 and non-reducing terminal linkages existed in polysaccharide P1A, but there are doubling amount of 1-->2 and 1-->4 linkages. Oxidable linkage of P1 A accounted for 45%, and inoxidable linkage of P1A accounted for 55%. Mannose, glucose and galactose were mainly linked by 1-->2 linkage. Rhamnose, arabinose and xylose were mainly linked by 1-->2 and 1-->4 linkages. PlA contained beta-Glc(1,6)-,beta-Gal(1,3)-,beta-Man(1,4)-beta-Rha,-Glc(1,4)-, Glc(1)-,-Gal(1,4)- and Man(1)-.
Acanthaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Molecular Weight ; Polysaccharides ; chemistry

OBJECTIVETo evaluate the effects of Icariin (ICA) on the proliferation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) in vitro and in vivo.

METHODSAn enzymatic digestion block was used in vitro to culture hPDLSCs, which were separated and purified by limited dilution cloning. The hPDLSCs were identified using cell-surface markers and cocultured with 1 x 10(-7) mol.L-1 ICA solution. The proliferation ability of these cells was determined by thiazolyl blue tetrazolium bromide (MTT) assay. After staining with alkaline phosphatase (ALP), osteogenesis was detected by enzyme-linked immunosorbent assay. Osteoblast-related genes were analyzed by reverse transcription-polymerase chain reaction. Alizarin red staining was performed to measure the level of calcium deposition. The hPDLSCs were cocultured with 1 x 10(-7) mol.L-1 ICA and nano-hydroxyapatite scaffolds in vivo before transplantation into subcutaneous tissues of nude mice. Osteogenic abilities were histochemically analyzed after 30 days of induction.

RESULTSThe hPDLSCs were affected by 1 x 10(-7) mol.L-1 ICA, and MTT assay showed that the proliferation of the groups treated with ICA in vitro was better than that of the control groups on the second day. The ALP activity of the treated hPDLSCs was significantly enhanced after cell culture for 3, 5, and 7 days. The gene expression of osteoblastic markers was also significantly enhanced after 7 days. The deposition of mineralization after incubation with 1 x 10(-7) mol.L-1 ICA increased compared with the control after cell culture for 14, 21, and 28 days. Furthermore, the bone expression of the treatment groups in vivo was significantly enhanced compared with that of the control groups.

CONCLUSIONTreatment with 1 x 10(-7) mol.L-1 ICA can significantly promote proliferation and differentiation of hPDLSCs in vitro and in vivo. ICA can effectively function as a bioactive growth factor in periodontal tissue engineering to replace traditional growth factors.

Alkaline Phosphatase ; Animals ; Cell Culture Techniques ; Cell Differentiation ; Cell Proliferation ; Coculture Techniques ; Enzyme-Linked Immunosorbent Assay ; Flavonoids ; Humans ; Mice ; Mice, Nude ; Osteoblasts ; Osteogenesis ; Periodontal Ligament ; Stem Cells

AIMTo investigate the effect of CSF contacting neurons (CSF-CNs) lesion in rat dorsal raphe nucleus (DRN) on the scores of morphine withdrawal symptoms precipitated by naloxone and the nNOS expression in dorsal horn of spinal cord, and study the relationship between the distal CSF-CNs in rat brain parenchyma and the development of morphine dependence and withdrawal.

METHODSChemical lesion of neurons the injection of cholera toxin subunit B with horseradish peroxidase (CB-HRP) into one of the rats lateral ventricles, TMBST reaction, nNOS immunohistochemistry and Western blot were used in this study.

RESULTSThe withdrawal symptoms by the naloxone precipitated attenuated obviously after the lesion of CSF-CNs in rat DRN, scores of all signs were significantly decreased about 38% compared to that of withdrawal group without lesion (P < 0.05). The withdrawal symptoms scores of vehicle withdrawal group and side lesion withdrawal group were not changed significantly (P > 0.05). Neurons in the location of CSF-CNs concentrated in the rat brain slices of lesion group were damaged obviously, there were only few CB-HRP positive neurons around the lesion location. But the location and the quantity of the CB-HRP positive neurons in the brain slices of the group without lesion was stable relatively, and their appearance was very clear. After the lesion, the nNOS expression and the quantity of the nNOS positive neurons in dorsal horn of spinal cord decreased significantly compared to that of withdrawal group without lesion (P < 0.05), but it also increased significantly compared to that of normal group and dependence group (P < 0.01).

CONCLUSIONThe lesion of distal CSF contacting neurons attenuated the scores of morphine withdrawal symptoms precipitated by naloxone and the nNOS expression in dorsal horn of spinal cord. The distal CSF contacting neurons in rat brain parenchyma partly participated in the development of morphine dependence and naloxone precipitated withdrawal possibly by the modulation of NO (nitric oxide).

Animals ; Brain ; drug effects ; pathology ; Male ; Morphine Dependence ; metabolism ; Neurons ; drug effects ; pathology ; Nitric Oxide Synthase Type I ; metabolism ; Raphe Nuclei ; cytology ; pathology ; Rats ; Rats, Sprague-Dawley ; Substance Withdrawal Syndrome ; metabolism