BACKGROUND: Embryonic stem cell (ESC) is a kind of highly undifferentiated totipotent cell. It can proliferate and maintain its totipotency in the system cultured in vitro. It is one of most promising stem cells in thetreatment of central nerve injury.OBJECTIVE: To observe the survival and migration of induced transplanted ESC in mice with spinal injury and hypoxic-ischemic encephalopathy.DESIGN: A completely randomized grouping design, controlled animal experiment.SETTING: Laboratory of Developmental Biology Research Center of Shanghai Second Medical University.MATERIALS: Sixty C57/BL6J mice, of clean grade and either gender, aged 6 to 8 weeks (n =30) and 7 days (n =30)were provided by the Shanghai Experimental Animal Center, Chinese Academy of Sciences [Permission No, SCXK (hu)2003-0003]. This animal experiment was approved by Animal Ethics Committee. Mouse ESC strain S8, labeled LacZ marker gene (Provided by Shanghai Developmental Biology Research Center). X-gal dyeing reagent (Sigma Company).METHODS: This experiment was carried out in the laboratory of Shanghai Developmental Biology Research Center (Shanghai Key Laboratory) from October 2002 to December 2003. ① Experimental grouping of spinal injury: Sixteen C57/BL6J successful mice models, aged 6-8 weeks, were randomized into 2 groups: experimental group (n =8), in which, following right spinal semi-sectioning, derivated cell suspension for inducing the in vitro differentiation of ESC was injected at 1 cm away from injury through vertebral canal, and control group (n =8), in which, following right spinal semi-sectioning, phosphate buffer solution (PBS) was injected at the peripheral region of injury. ② Hypoxic-ischemic encephalopathy experimental grouping: Sixteen successful C57/BL6J mice models, aged 7 days, were randomized into 2 groups: experimental group (n =8), following ligation of right common carotid artery, mice were placed in the closed container containing 0.08 volume fraction of oxygen and 0.92 volume fraction of Nitrogen gas, and taken out 1.5 hours later; 3 μL ESCs were injected into the right cerebral ventricle at about 1 week, and control group (n =8), in which, the same amount of PBS was injected into the right cerebral ventricle. ③ At 12 weeks after transplantation, the survival and migration of induced ESCs labeled by Lac-Z in the spinal cord and brain were observed by zymologic method.MATN OUTCOME MEASURES: Survival and migration of ESCs in the central nervous system.RESULTS: ①After being induced in vitro and transplanted to spinal injured region, ESCs differentiated into neural precursor cells. Neural precursor cells could survive in the injured region and migrate to 5 mm away from injured region.Immunohistochemistry proved that the neural precursor cells of transplanted ESCs could differentiate into neurons.Morphologically, it was proved that neural precursor cells-derived from ESCs could well integrate peripheral tissue. ② The induced ESCs were injected into the lateral cerebral ventricle of mice. Derived ESCs widely distributed in the injured hippocampal region, cerebral cortex ventricle choroid plexus, vascular endothelium and other regions, and integrated peripheral tissue, which were similar to adjacent cells in morphology, suggesting that induced ESCs also could survive for long time and far migrate.CONCLUSION:The induced ESC can survive and migrate in the host injured brain and spinal cord, and the migration of ESCs is more obvious in the brain than in the spinal cord.

Objective To investigate the repair and protective effect of extracorporeal membrane oxygenation (ECMO) on liver and bile duct after cardiac death in pig.Methods Eight pigs were purchased and cardiac arrest was induced by the administration of 1 g KCL intravenously,followed by 30 min cardiopulmonary resuscitation according to standard guideline.Cannulas were placed through inferior vena cava and abdominal aorta,and then connected to ECMO extracorporeal circulation pipes.ECMO was performed for 4 h.Circulation flow rate of hepatic artery and bile production were monitored and recorded.Lactate dehydrogenase (LDH),γ-glutamyl transferase (γ-GT) and direct bilirubin (DBIL) in bile were detected.Transaminase,tumor necrosis factor-α (TNF-α),interleukin-1β (IL-13),hyaluronic acid (HA),endothelin-1 (ET-1) and nitric oxide (NO) in serum were detected.Pathological change was observed by HE staining under optical microscope and cell apoptosis was detected by TUNEL.Results There was no bile production after cardiac death,which increased to 80％ of the baseline after 4h of ECMO.In addition,γ-GT,LDH and DBIL content in bile was (23.3 ± 11.8) IU/L,(15.9 ± 3.3) IU/L and (72.3 ± 21.4) mmol/ L,and IL-1,TNF-α and HA content in serum was (117.6 ± 39.0) ng/L,(120.4 ± 16.5) ng/L and (63.7 ± 4.4) ng/L,respectively,and no statistically significant differences were observed when compared with the baseline (all P ＞ 0.05).ET-1 content was (4.9 ± 1.3) ng/L and NO content was (135.3 ± 16.7)mmol/L in serum,which was statistically increased (both P ＜ 0.05).Pathological changes of liver and bile duct were significantly alleviated.Conclusion ECMO could exert protective effect on liver and bile duct after cardiac death.

ObjectiveTo explore the effect of psychological intervention on anxiety of patients with motor neuron disease (MND) before stem cell transplantation via lumbar puncture.Methods40 MND patients were randomly divided into the group A and group B with 20 cases in each group. Every patient received anxiety assessment on the day of hospitalization, the day before transplantation as well as the day after transplantation. As to the patients in the group B, psychological intervention was performed before the second time anxiety assessment, while no such intervention to the ones in the group A. Then the assessment was statistically analyzed.ResultsThe patients in each group had anxiety. After psychological intervention, the assessments of the second and the third time of the patients in the group B were better than that of the patients in the group A ( P<0.01).ConclusionPsychological intervention before transplantation can effectively ease the anxiety of patients.

We investigated the pathogen of an outbreak of lung infection with unknown causes.By epidemiological analysis,we used real-time PCR,ELISA,gold dipstick,VITEK2 and MALDI-TOF-MS to identify suspicious bacteria.We made use of serum plate agglutination test to confirm the suspicious bacteria and the patient serum.We isolated 2 strains of Cryptococcus albidus from environmental samples.There has been specific agglutination between suspicious bacteria and patient serum.This pneumonia may be related to the infection of Ccryptococcus albidus.

OBJECTIVETo study the influence of repeated seizures and anti-epileptic drug on phosphorylated cAMP response element binding protein (pCREB) in rat model of cognitive impairment, and the effect of Caoguo Zhimu Decoction (CZD) on it.

METHODSOn the basis of epileptic model induced by pentylenetetrazol (PTZ), cognitive impairment model was induced by kindling epilepsy with PTZ everyday, which were then di-vided into the model-1 group, the CZD-1 group, the nimodipine-1 group, and those by injecting large dosage phenytoin sodium were divided into the model-2 group, the CZD-2 group and the nimodipine-2 group. Changes of pCREB protein in rat's hippocampus were detected using immunohistochemistry and Western blotting assay.

RESULTSThe expression of pCREB was higher in the CZD-1 and nimodipine-1 group than in the model group, while it was significantly different in the CZD-2 and nimodipine-2 groups than in the model-2 group.

CONCLUSIONCZD could relieve the cognitive dysfunction in the epileptic model rats induced by everyday PTZ kindling or by dilantin through increasing the pCREB expression.

Animals ; Cognition Disorders ; chemically induced ; prevention & control ; Cyclic AMP Response Element-Binding Protein ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Epilepsy ; chemically induced ; drug therapy ; metabolism ; Hippocampus ; drug effects ; metabolism ; pathology ; Kindling, Neurologic ; drug effects ; Male ; Pentylenetetrazole ; Phenytoin ; adverse effects ; Phosphorylation ; Phytotherapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley

The chemical consitituents from cytotoxic fraction of the Callicarpa nudiflora extract were isolated and purified by a combination of HP-20 macroporous resin, silica gel and Sephadex LH-20 column chromatographies. The structures were elucidated on the basis of the spectroscopic data and comparison of their spectroscopic data with reported data. The cytotoxicity was evaluated by the MTT assay. The 50% and 70% EtOH elutions of EtOH-extract showed significant cytotoxic activities, leading to the isolation of twelve compounds, which were identified as luteoloside(1), lutedin-4'-O-β-D-glucoside(2), 6-hydroxyluteolin-7-O-β-glucoside(3), lutedin-7-O-neohesperidoside(4), rhoifolin (5), luteolin-7, 4'-di-O-glucoside (6), forsythoside B (7), acteoside (8), alyssonoside (9), catalpol(10), nudifloside(11), and leonuride(12). Compounds 3-6, 10 and 12 were isolated from this genus for the first time, and compound 9 was isolated from this plant for the first time. The cytotoxicity assay demonstrated that flavonoids 1-6, in various concentrations, showed monolithic proliferation inhibitory activities against Hela, A549 and MCF-7 cell lines. Compounds 3, 5 and iridoid glycoside 11 possessed higher cytotoxicacivities. In short, flavonoids are the main components of cytotoxic extract from C. nudiflora, while phenylethanoid glycosides are the predominant ingredient but inactive to cancer cell lines. In addition, the minor iridoid glycoside expressed weak cytotoxic activity.
Callicarpa ; chemistry ; Cell Proliferation ; drug effects ; Cytotoxins ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Humans ; MCF-7 Cells ; Molecular Structure

Objective The nano-zirconia scaffolds we previously prepared had a good 3-dimensional ( 3D ) connectivity but did not achieve the ideal sintering rate and compressive strength .The objective of this study was to explore the enhancing effect of polyvinyl butyral ( PVB) as a dispersant on the compressive strength of 3D nano-zirconia porous scaffolds for bone tissue engineering . Methods We prepared the slurry containing different concentrations of PVB and ana-lyzed the improving effect of PVB on the mechanical properties of the scaffolds by sediment experiment , compressive strength test and scan-ning electron microscopy . Results The sediment experiment showed
no significant stratification in the slurry with 0.2wt%PVB, white suspension in the upper layer and white precipitate in the lower layer , with a significantly higher compressive strength of the scaffold ([0.324 ±0.030] MPa) than that of the scaffold prepared by adding other concentrations of PVB to the slurry (P <0.01).And the compressive strength of the scaffold constructed by adding no dispersant ([0.109 ±0.021] MPa) was remarkably lower than that of the scaffold constructed by adding PVB to the slurry (P<0.05).Scanning electron microscopy demonstrated that the scaffold prepared by adding 0.2wt%PVB to the slurry had a complete porous structure with the fewest and most sparsely distributed surface cracks as compared with other PVB concentration groups . Conclusion PVB can signifi-cantly improve the stability of zirconia slurry , enhance the compressive strength of the nano-zirconia porous scaffold , and make the scaf-fold more applicable to bone tissue engineering .

Objective To investigate the effect of calpeptin on diaphragmatic injury and atrophy under controlled mechanical ventilation in rats.Methods A total of 24 SPF Sprague-Dawley (SD) rats were randomly divided into anesthetized control group (CON group),24-hour controlled mechanical ventilation group (CMV group),and 24-hour CMV + treatment with calpeptin group (CMVC group),with 8 rats in each group.Animals in the CON group received an intraperitoneal injection of pentobarbital sodium without CMV and continuous infusion of pentobarbital sodium.A small-animal ventilator was used for 24 hours in rats of CMV group.Rats of CMVC were treated with a specific calpain inhibitor calpeptin (4 mg/kg).The drug was injected subcutaneously 2 hours before and 8,15 and 23 hours after mechanical ventilation.Changes in diaphragm ultrastructure,light microscopic picture,and myosin heavy chain (MHC) expression were observed.Results ① Alignment of myofilaments and normal Z-band,and the shape of mitochondria were maintained in CON group as revealed by electron microscope.The signs of misalignment of myofibrils,disruption of Z-band and vacuolar mitochondria were found in CMV group,and they were obviously improved in CMVC group.The density of muscle injury (× 10-2/μm2) in CMV group was significantly higher than that in control group (36.8 ± 13.7 vs.6.4 ± 6.3,t=6.373,P=0.001),and that in CMVC group was significantly lowered (17.6 ± 9.1 vs.36.8 ± 13.7,t=3.694,P=0.002).② In CON group,the diaphragm fibers appeared regular in cross section without pathologic change under light microscopy.Fuzzy muscle striations,irregular muscle fibers,centralized nuclei and swelling of capillary endothelial cells were observed in CMV group,while pathological changes in the CMVC group were milder significantly.③ In CMV group,the density of MHCslow and MHCfast was lower compared with that of CON group,and the gray value was lowered by 61.1％ (t=8.138,P=0.001) and 77.1％ (t=8.844,P=0.001),respectively,especially in MHCfast.However,the gray values of MHCslow and MHCfast were increased by 1.51 folds (t=4.601,P=0.010),and 1.33 folds (t=2.859,P=0.011),respectively,after treatment with calpeptin,and the elevation was more significantly in MHCslow.Conclusions Diaphragmatic injury and atrophy were found after CMV for 24 hour.Calpeptin could reverse the detrimental effects of CMV,and it suggested that calpain plays an important role in modulating the ventilator-induced dysfunction of the diaphragm.

Objective To investigate the presence and significance of Th9 cells and the related transcription factor ( PU.1 ) and cytokine ( IL-9 ) in peripheral blood of patents with hashimoto thyroiditis ( HT) .Methods Thirty patients with HT and thirty age/gender matched healthy subjects were recruited in this study.The peripheral blood and serum samples were collected from each subject.The percentages of Th9 cells and the transcriptional levels of PU.1 in peripheral blood mononuclear cells ( PBMCs) were meas-ured by flow cytometry analysis and real-time RT-PCR.The concentrations of IL-9, the functions of thyroid and the titers of thyroid-specific autoantibodies ( TPOAb and TgAb) in serum samples were detected by using enzyme-linked immunosorbent assay ( ELISA ) and electrochemiluminescence immunoassay analysis ( ECLIA) .Results Compared with healthy subjects, the percentages of Th9 cells and the expression of PU.1 at mRNA level in PBMCs and the concentrations of IL-9 in serum samples were all significantly in-creased in patients with HT [(1.49±0.68)%vs (0.87±0.24)%], 4.91±2.14 vs 1.66±0.52, (26.90± 7.74) pg/ml vs (16.71±5.87) pg/ml, all P<0.01).Serum concentrations of IL-9 were positively correla-ted with the percentages of Th9 cells (r=0.419, P=0.021).Moreover, the percentages of Th9 cells were positively correlated with the titers of TPOAb and TgAb in serum samples (r=0.394, P=0.032;r=0.457, P=0.011) of patients with HT.Conclusion The levels of Th9 cells and the related cytokine IL-9 were in-creased in the peripheral blood of patients with HT.A positive correlation was found between the percentage of Th9 cells and the titers of thyroid-specific autoantibodies.This study indicated that Th9 cells might be in-volved in the pathogenesis of autoimmune damage in thyroid.

Objective To investigate Feining granule on prevention and treatment of rat with radiation-induced lung injur ( RILI) and its effect on cytokine transforming growth factor-β1(TGF-β1), interleukin-1 (IL-1), interleukin-6 (IL-6).Methods 105 SD female rats were selected, according to random number table, and divided into seven groups: normal group, model group, dexamethasone group(positive drug), Feining granule low-dose group (FN low-dose group), FN medial-dose group, FN high-dose group and FN prevention group, 15 rats in each group.Except for normal group, all remaining groups received the X-ray irradiation of 15Gy, DT30Gy/2f/1w.FN prevention group were intragastric infused with FN granule one week before irradiation, and the other groups 48 h after irradiation.Five rats were sacrificed randomly at 2, 4, 6 weeks respectively, and right lung tissues were taken out.The contents of TGF-β1, IL-1 and IL-6 were detected by immunohistochemical method.ResuIts TGF-β1, IL-1 and IL-6 contents in lung tissue of model group at 2, 4, 6 weeks were higher than those of normal group (P<0.05).The above indicators after treated by Feining granule were lower than those of model group at each time (P<0.05),with a concentration-dependence manner to some extent.The above indicators in FN high-, medial-and low-dose group were higher than those in dexamethasone group (P<0.05), to some extent.However, the above indicators in FN prevention group were lower than those in dexamethasone group ( P<0.05 ) .ConcIusion Feining granule could prevent and cure radiation induced lung injury through decreasing TGF-β1, IL-1 and IL-6 content.The efficacy of dexamethasone is stronger than FN treatment groups, but is weaker than FN prevention.