Objective To investigate the effect of hyperoxia on the transdifferentiation level of type Ⅱ alveolar epithelial cells (AEC Ⅱ) in vivo and in vitro,in order to illuminate the mechanism of epithelial injury in bronchopulmonary dysplasia (BPD).Methods Newborn Wistar rats were randomly devided into control group (room air inhalation) or model group (85％ oxygen inhalation) after birth.Lung tissue sampling and AEC Ⅱ isolation was conducted on 7 d,14 d,21 d.Type Ⅰ alveolar epithelial cells (AEC Ⅰ) marker aquaporin 5 (AQP5) and AEC Ⅱ marker surfactant protein C (SP-C) were examined by Western blot and florescent real-time PCR.AEC Ⅱ isolated from normal newborn rats was randomly devided into normoxia group (21％ oxygen) or hyperoxia group (85％ oxygen) after 24 h culture,and continued culturing for another 48 h in vitro.Then the morphological changes of cells were observed under inverted phase contrast microscope.The expression and location of markers for AEC Ⅰ and AEC Ⅱ was examined by immunofluorescence double staining.The protein expression of AQP5 and SP-C was evaluated by Western blot,and the mRNA expression of these markers was examined by florescent real-time PCR.Results In AEC Ⅱ isolated from the animal models,the AQP5 protein expression increased from 7 d while the SP-C expression decreased from 14 d in the model group comparing with the control group.In the model group,AQP5 mRNA expression increased and SP-C mRNA expression decreased since 7 d after hyperoxia exposure (P ＜ 0.05),with the difference between groups more obvious as exposure time extending.After culturing in vitro,AEC Ⅱ isolated from normal newborn rats expressed more AQP5 and less SP-C,with more cells double stained in the hyperoxia group compared with the normoxia group,examined by immunofluorescence double staining.The protein and mRNA examination results both showed that AQP5 expression increased and SP-C expression decreased in the hyperoxia group compared with the normoxia group (P ＜0.01).Conclusion After hyperoxia exposure,no matter in vivo or in vitro,the expression of AEC Ⅱ marker SP-C decreases while the expression of AEC Ⅰ marker AQP5 increases.These results indicate that the excessive transdifferentiation of AEC Ⅱ takes part in the recovery process after hyperoxia induced lung injury.

Objective To investigate the variation in expression and the significance of markers indi-cating typeⅠalveolar epithelial cells ( AECⅠ) and type Ⅱ alveolar epithelial cells ( AECⅡ) in hyperoxia induced bronchopulmonary dysplasia( BPD) model. Methods A total of 80 term normal Wistar rats were randomly devided into model group (85% oxygen) or control group (room air) within 12 h after birth,with 40 rats in each group. On day 7,day 14,day 21 after exposure,the pathological characteristics of lung tissues were observed using HE staining, the expression and location of AECⅠ marker aquaporin 5 ( AQP5 ) and AECⅡmarker surfactant protein-C ( SP-C) were examined using immunofluorescence double staining. West-ern blot analysis was employed to examine the expressions levels of AQP5 and SP-C proteins,while real-time PCR was used to evaluate the mRNA expression of AQP5 and SP-C. Results Alveolar developmental disor-der was observed in lung tissues of the model group,including fewer,larger,simplified alveoli,thicker alveo-lar walls,and fewer alveolar secondary septa. Immunofluorescence double staining showed increased and dis-organized AQP5 and SP-C expression, with significantly higher ratio of double-stained cells/SP-C positive cells in the model group ( P<0. 001 ) . Comparing to the control group, the expression of AQP5 and SP-C protein increased from 7 d after hyperoxia exposure,which continued to 21 d. The mRNA expression levels of these two markers both significantly increased in the model group compared with the control group, with AQP5 starting from 7 d while SP-C starting from 14 d after hyperoxia exposure (P<0. 05),and the differ-ence between two groups became more significant with the exposure time extending. Conclusion The expression of AECⅠ marker AQP5 and AECⅡ marker SP-C both increase in the lung tissues of hyperoxia induced BPD newborn rats,with more AECⅡ transdifferentiated into AECⅠ. These changes of the markers indicate that there is excessive transdifferentiation of AECⅡ in the recovery process after BPD lung injury.

Objective To investigate the ultrastructural alteration in brain tissues as well as the expression of bone morphogenetic protein(BMP) 4 and its effects on regulating myelination in the process of white matter injury development.Methods A total of 152 Sprague-Dawley newborn rats(3 days old) were randomly divided into white matter injury group(n=76) or control group(n=76).The white matter injury model was established by ligation of the right common carotid artery and hypoxic exposure(8% O2 and 92%N2),and samples were collected at 3d,7d,14d and 21d after operation.Morphological changes of the brain tissues were observed under a light microscope,while myelination was analyzed using a transmission electron microscope.The expression and location of BMP4 and myelin basic protein(MBP),a marker for myelination,was detected by immunohistochemistry staining,expression levels of BMP4 and MBP proteins were analyzed by Western blotting,and BMP4 mRNA expression was measured by real-time PCR.Results Observed under the light microscope,the cellular structure was clear,fibers arranged closely and orderly in the white matter of the control group.Whereas in the white matter injury group,sparse cells,loose mesh shaped white matter,and disorderly oriented fibers were observed.In the control group,myelin sheath had regular morphology,uniform density,and same thickness,observed using the transmission electron microscope.While in the white matter injury group,the myelin sheath was loosened,thinned,lamellar separated,and boundary obscured.Using immunohistochemistry staining,Western blot,and real-time PCR analyses,it was found that the protein and mRNA expression of BMP4 had no significant change with the increase of age in the control group,while it was rapidly increased with the extending of ischemic time in the white matter injury group.Comparing with the control group,the expression of BMP4 was significantly increased since 3d after operation in the white matter injury group(P<0.05),and the difference between two groups became more significant with the extending of ischemic time.The expression of MBP protein was analyzed by immunohistochemistry staining and Western blot,and a gradual increase was found in both groups with the increase of age.However,the expression of MBP protein was significantly decreased on 14d and 21d after operation in the white matter injury group compared with the control group(P<0.05).Conclusion Myelination disorders exists in white matter injury induced by ischemia-anoxemia.Meanwhile,the expression of BMP4 is significantly increased in the white matter injury group,indicating a possibility that BMP4 involves in the regulation of myelination disorders in white matter injury.

In this paper,a unique case of organ transplantation was inspected by organ transplant ethics committee,through which we try to investigate the mode of current operation,problems and confusion of organ transplant ethics committee in China by the constitutive principles,inspection scope,process,content,especially the functional authority and other relevant contents of hospital ethics committee.

This paper describes the role,method and experience,and working model of medical ethics in large-scale hospitals,and holds that the modern hospital management mode has transformed from scientific management to humanism-based management,whose essence is the transformation of managing concept,and the embody of management ethics and management culture.Important connotations and primary task including employing virtue in hospital management,obeying legal regulations in medical practice,and conducting humanistic medicine should be emphasized and concerned in hospital management.

AIM: To investigate the effect of ethanol and chemical carcinogen N,N'-dinitrosopiperazine(DNP) on the exogenous expression of CYP2E1.METHODS: Exogenous hCYP2E1 was introduced into NIH3T3 mediated by lipofectamine. Then the integration of exogenous gene was showed by Southemrn blot. After treated with different concentration of ethanol and DNP,RT-PCR and Western blotting were used to analyze the expression change of hCYP2E1 in NIH 3T3. RESULTS: Two cell clones with integration and stable expression of exogenous hCYP2E1 were obtained. The RT-PCR and Western blotting showed that human CYP2E1 mRNA and protein expression was enhanced with increase in ethanol and DNP concentration. CONCLUSION: Exogenous expression of hCYP2E1 was steadily induced by ethanol and DNP. The mechanism may be due to the activation of its transcription. The DNP carcinogenesis might be related to its in situ activation by CYP2E1.

AIM:To observe the effects of hawthorn leaf polymeric procyanidins ( PPC) on calcium mobiliza-tion of vascular endothelial cells , and to study the underlying mechanism .METHODS: Free calcium in cultured human umbilical vein endothelial cells (HUVECs) was labeled with Fura-2.HUVECs were treated with ATP, a positive control drug, and PPC at concentrations of 12.5, 25 and 50 mg/L..The intracellular calcium concentrations were measured with a living cell microscope for 30 min.RESULTS:PPC concentration-dependently increased the intracellular calcium concen-tration of HUVECs .The intracellular calcium concentrations in 25 and 50 mg/L PPC groups were significantly higher than that in normal group (P<0.01).The dynamic manner of calcium concentration elevations elicited by PPC was a slow in -crease which happened after a latency time of several minutes , lasted for several minutes , and reached a plateau finally . This manner was quite different from that elicited by ATP , a typical SOC operator , hinting different mechanisms between them .Inhibiting the intracellular calcium release did not influence the effects of PPC;however , deleting extracellular calci-um, inhibiting the reverse mode of Na +-Ca2+exchange, or deleting extracellular sodium , restrained or even abolished the effects of PPC.CONCLUSION:PPC elicits calcium mobilization in vascular endothelial cells , which may be one of the mechanisms of the vascular modulatory activity of hawthorn procyanidins .This effect may be achieved through inducing the influx of sodium and then activating the reverse mode of Na +-Ca2+exchange.

BACKGROUND: Human adipose tissue-derived stromal cells (ADSCs) possess exert proliferation and multi-directional ability. As a novel stem cell, it has been widely utilized in tissue engineering and plays an important role in biological and potential therapeutic applications.OBJECTIVE: To review the research progress, applications and existing problems of human ADSCs in tissue engineering and cell therapy by retrieving relevant publications. METHODS: PubMed and CNKI databases were undertaken to identify the relevant articles published from January 1960 to January 2009 with the key words of "human adipose tissue-derived mesenchymal cells, isolation, differentiation, immune phenotype, application" both in English and Chinese. The articles relate to biological characteristics and applications of ADSCs were selected. In the same field, the documents published earlier or in the authoritative journals were preferred, and the repetitive studies were excluded. RESULTS AND CONCLUSION: A total of 81 documents were searched by computer, and 57 references were included in the final analysis. The results demonstrated that, human ADSCs share many characteristics, including the high proliferative potential and exhibiting the exert ability to undergo multilineage differentiation under appropriate conditions. Human ADSCs can not only utilize for tissue repairing, but also for cell immune modulation and gene therapy. However, there are still problems in its application. With the development of research on human ADSCs, their biological characteristics will be revealed, and their application in tissue repairing, cell therapy, transplantation, as well as gene therapy must be hold a great promise.

Objective To clinically analyze the incidence of early extrapulmonary complications in premature infants with bronchopulmonary dysplasia(BPD),including periventricular intraventricular hemorrhage(PVH-IVH),white matter injury(WMI),parenteral nutrition associated cholestasis(PNAC) and metabolic bone disease(MBD),in order to direct the prevention and monitoring of these complications in BPD patients.Methods The clinical data of premature infants who were admitted to the neonatal department between September 2014 and December 2015 was retrospectively analyzed.A total of 87 premature infants diagnosed with BPD were studied as BPD group,while other 90 premature infants without BPD who were hospitalized at the same time were randomly selected as non BPD group.The occurrence of several common extrapulmonary complications was compared between two groups,including PVH-IVH,WMI,PNAC and MBD.Results The incidence of PVH-IVH in BPD group increased compared with non BPD group[(26.4%(23/87) vs 11.1%(10/90)] (P＜0.01),grade Ⅰ-Ⅱ PVH-IVH was more often seen in the BPD group too[24.1%(21/87) vs.11.1%(10/90)](P＜0.05),although the difference between two groups regarding the incidence of grade Ⅲ-Ⅳ PVH-IVH was not significant (P＞0.05).The incidence of WMI in BPD group was much higher than that in non BPD group[33.3%(29/87) vs 16.7%(15/90)] (P＜0.05),especially periventricular leukomalacia,the severe type of WMI,was more often found in BPD group than that in non BPD group[13.7%(12/87) vs 2.2%(2/90)](P＜0.05).The incidences of PNAC[22.9%(20/87) vs 5.5%(5/90)],MBD[17.2%(15/87) vs 3.3%(3/90)] and MBD with imaging changes[6.9%(6/87) vs 0] were all higher in BPD group compared with non BPD group,with significant differences between the two groups (P＜0.05).Conclusion BPD patients are more likely to have early extrapulmonary complications like PVH-IVH,WMI,PNAC and MBD than other preterm infants.It is crucial to prevent these complications reasonably and monitor them regularly for the BPD patients in order to improve the quality of life.

Objective To detect the proliferation and the expression levels of downstream target genes of Notch pathway of T-ALL CCRF-CEM(CEM)cell line treated with exogenous DLK1 protein,in order to investigatethe effects of DLK1 protein on the Notch pathway in CEM cells.Methods CCK-8 assay was performed to examine the proliferation of CCRF-CEM cells,which were treated with various concentration(0.5,1.0,1.5 μg/ml)DLK1 for various time(24,48,72 h).RFQ-PCR was applied to assess the mRNA expression level of Notch1 receptor and downstream target genes of Notch pathway in CEM cells,which were treated for various time(24,48,72 h).Results DLK1 protein stimulated the proliferation of CCRF-CEM cells,and the proliferation rates with different concentrations of DLK1 were 0.14±0.03,0.17±0.04,0.55±0.01 in 72 hours,respectively,there were statistic differences between that in the experimental group and that in the control group(P＜0.05).DLK1 protein up-regulated the Notch1 receptor and its downstream target genes HES1,c-myc and NF-κB.The relative transcript levels of target genes HES1 in 72 hours,c-myc in 48 hours and NF-κB in 72 hours were 3.2551±0.3100,1.6086±0.0941,2.0515±0.3453 respectively,and there were statistic differences between that in the experimental group and that in the control group(P＜0.05).Conclusion DLK1 protein stimulates the proliferation of T-ALL CCRF-CEM cells by up-regulating Notch1 receptor and downstream target genes HES1,c-myc and NF-κB of Notch pathway.