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.

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 To investigate methods and experience of autologous granular fat transplantation by correction of aging face. Methods 178 cases of aging face subjects were treated by liposuction. Then autologous granular fat was harvested by liposuction,centrifuged and purified, and injected into marked areas of aging face. The injection process should be controled by multilevel and multiple tennel, in order to supplement the loss of facial soft tissue volume and improve the aging face. Results The follow-up period ranged from 3 months to 3 years. All the subjects had a satisfactory results with no obvious complications such as fat liquefaction, infection and ulceration. The effect of autologous granular fat transplantation was obvious and permanent. Conclusions Autologous granular fat transplantation is a safe and effective surgical technique for facial soft tissue augmentation that can effectively improve aging face and worthy of clinical application.

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 evaluate the effect of online teaching platform in clinical neurology prac-tice. Methods The participants were medical undergraduate students in the process of clinical practice in Department of Neurology of XiJing hospital, who were divided into the traditional practice teaching group (n=77) and online platform auxiliary practice teaching group (n=90), respectively carry-ing out the corre-sponding rotation practice teaching . Unified objective structured clinical examination was given to two groups of students to implement departmental rotation examination , and investigation was made among teachers and students on the evaluation of the effect of practice. SPSS 19.0 was used to do t test to the two sets of corresponding evaluation data. Results Total score of departmental rotation examination in online platform auxiliary practice teaching group (69.33 ±2.74) was significantly higher than traditional teaching group (67.23±2.50) (P=0.000). Scores of basic theories, skill and operation, clinical case analysis in online platform auxiliary practice teaching group were significantly higher than traditional teaching group (P<0.05). However, medical record writing scores in online platform auxiliary practice teaching group (7.39± 1.09) were significantly lower than the traditional teaching group (8.03±1.03) (P=0.000). Teachers' eval-uation of practice effect was obviously higher in online platform auxiliary practice teaching group (12.33± 0.52) than that in traditional teaching group (10.67 ±1.03) (P=0.005). Students' evaluation of interest in learning, content arrangement, teachers' sense of responsibility, evaluation of teaching ability, practice effect were obviously higher than that of traditional teaching group (P<0.05). Conclusion This study preliminary has achieved good practical effect by using online auxiliary teaching platform, and got high evaluation by both teachers and students. Inter-active communication and sharing of resources is easy in the online plat-form of practice teaching . On-line platform will help students develop with clinical neurology diagnosis thinking.

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 optimal time of extracorporeal membrane oxygenation (ECMO) on repairing Maastricht Ⅱ pig liver with warm ischemia injury for 30 min.Method Thirty-six miniature pigs were randomized to ECMO 2-h group,ECMO 4-h group and ECMO 6-h group,12 pigs per group,6 donors and 6 recipients.Cardiac arrest was induced by administration of 1 g KCl intravenously,followed by cardiopulmonary resuscitation (CPR) for 30 min according to the standard guideline.Cannulas was placed through inferior vena cava and abdominal aorta,then connected to ECMO extracorporeal circulation pipes.Transaminase,circulation flow rate of portal vein and hepatic artery and arterial blood gas were monitored and recorded.The hepatic tissues were cut into sections for pathological observation by HE stain under a light microscope.Apoptosis was detected by TUNEL and apoptotic index was calculated.The liver was stored in cold UW for 2 h after the ECMO circulation,then orthotopic liver transplantation without veno-venous bypass was performed.The levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured in the peripheral blood for 5 days after the operation.Result With the increase of the running time of ECMO,transaminase and lactate levels were decreased continuously.Circulation flow rate of portal vein and hepatic artery in ECMO 6-h group was significantly lower than that in the other two groups (P＜0.05).Pathological change in ECMO 4-h group was milder than the rest two groups.Apoptosis index in ECMO 2-h,4-h and 6-h groups was (40.20 ± 7.22)％,(18.60 ± 4.04)％ and (29.25 ± 5.98) ％,respectively.The 5-day suvival rate in ECMO 2-h,4-h and 6-h groups was 83％,100％ and 83％,respectively.The transaminase level in ECMO 4-h group at 5th day after the operation was lower than in ECMO 2-h group and 6-h group (P＜0.05).Conclusion The optimal time of ECMO on repairing Maastricht Ⅱ liver was 4 h.The effect of restoration is not ideal when circulation time is not enough.Liver function and liver cell viability decline beyond 4 h.