Background:Disruption of intestinal epithelial tight junction and the followed barrier function play important roles in the pathogenesis of intestinal disorders. Curcumin could provide protection for the impaired barrier function. Aims:To investigate the protective effect of curcumin on ethanol-induced intestinal mucosal barrier disruption. Methods:Caco-2 cells were cultured to establish intestinal epithelial cell barrier model in vitro,and then were divided into control group, ethanol group and different concentrations of curcumin groups(5,20,80 μmol/ L curcumin). Trans-epithelial electrical resistance(TEER)and flux of sodium fluorescein for Caco-2 cell monolayers were measured to examine intestinal epithelial barrier function. Expression and localization of Occludin protein were measured by Western blotting and immunofluorescence,respectively. Cell structure was observed by transmission electron microscopy( TEM). Results:Compared with control group,TEER was significantly decreased and flux of sodium fluorescein was significantly increased (P < 0. 05),expression of Occludin protein was significantly decreased(P < 0. 05)in ethanol group. Immunofluorescence showed that Occludin protein expression was discontinuous and fluorescence intensity was low. TEM showed that brusher border was disorganized,and cell-cell junction was vague. When pretreated with curcumin,the above-mentioned indices were significantly improved,especially in 20 μmol/ L curcumin group( P < 0. 05). Conclusions:Curcumin protects ethanol-induced intestinal epithelial cell barrier disruption.

Hematopoietic Stem Cell Transplantation ; Histocompatibility Antigens Class I ; genetics ; Humans ; Polymorphism, Genetic ; Transplantation, Homologous

Objective:To prepare and test tetrameric sH-2K~d-HBc complex for the further measurement of the specific CTL response.Methods:PE labled streptavidin with 4 biotinylated binding sites can bind to 4 biotinylated monomer to form the corresponding tetramer.Mice were immunized via different methods of genetic immunization by use of the construted pcDNA3-C plasmid to get the specific CTLs.Then our prepared tetramer was applied to stain the specific CTLs by the analysis of flow cytometry.Results:We applied our prepared tetramer to stain the cells from the experimental groups and control group.The results showed the tetramer was able to discriminate the frequencies of specific CTL induced by the three immunol methods(0.24%,0.26%,0.36% vs 0.07%,P≤0.05).This demonstrated that the prepared tetramer could bind its targets specifically and efficiently.The three immunol methods induced different levels of immune responses.Compared with the traditional muscle injection,gene gun induced weaker humoral immune response and stronger cellular immune response,and hydrodynamic injection induced the strongest humoral and cellular immune responses.Conclusion:Have successfully constructed the sH-2K~d-HBc tetramer.The techniques and methods can be used for preparation of tetramers of other types of MHCⅠ molecules.

Objective To determine the correlation between high serum levels of homocysteine (Hcy), low folate and
vitamin B12 levels and the clinical index of neonatal hypoxic ischemic encephalopathy (HIE), providing the reference for the diagnosis and treatment of neonatal patients with HIE. Methods Totally 94 cases of neonatal patients with HIE (32 mild, 40 moderate, and 22 severe) treated by Tianjin children’s hospital from April 2010 to April 2011 were enrolled. The control group was 20 cases of neonatal patients with neonatal swallowing syndrome at the same period. The serum levels of Hcy, folate and vitamin B12 were compared between four groups. Results There were significant differences in serum levels of Hcy, folate and vitamin B12 between four groups (P<0.05). Compared with the control group, there was higher level of Hcy and lower levels of folate and vitamin B12 in HIE groups. The Hcy levels increased with the deterioration of HIE in three HIE groups. The serum level of vitamin B12 was significantly lower in severe group than that in moderate group (P<0.05). Conclusion The serum Hcy level has certain reference value in tassessment of the severity of HIE. The supplementation of folate and vitamin B12 may play an active role in the process of the treatment in neonatal patients with HIE.

Objective To investigate the prenatal diagnosis and prenatal genetic conselling of pseudomosaic trisomy 20. Methods One case of pseudomosaic trisomy 20 was analyzed and relative literatures were reviewed. Results A 31-year-old gravid 1, para 0 woman underwent amniocentesis at 18 weeks of gestation due to high risk of trisomy 21 during maternal serum screening in September, 2012. Interphase fluorescence in situ hybridization (FISH) of amniocytes with probes GLP13/GLP21/CSP18/CSPX/CSPY showed a normal result, while cytogenetic analysis of cultured amniocytes revealed a karyotype of 47,XY,+20[7]/46,XY[9]. The level of trisomy in the cultured amniocytes was 7/16. Cordocentesis revealed a karyotype of 46,XY in cultured cord blood cells. Interphase FISH analysis was performed using the probes D20Z1 (20p11.1-q11.1) and D20S1157/20QTEL14 (20 per/qter). Each probe showed two signals in all uncultured amniocytes. The prenatal ultrasound findings were unremarkable. The mosaicism was considered to be pseudomosaicism. After genetic counseling, the parents selected to continue the pregnancy. A healthy male baby was delivered at 39 weeks of gestation. Postnatal cytogenetic analysis revealed a karyotype of 46,XY in peripheral blood lymphocytes. Interphase FISH analysis of the uncultured buccal cast-off cells using the probes D20Z1 and D20S1157/20QTEL14 showed normal results in 100%cells. There was no phenotypic abnormality at the age of seven months. Conclusions When mosaic trisomy 20 is identified in amniocytes, further evaluation and genetic counseling are required. Interphase FISH of the uncultured amniocytes with a chromosome-specific probe is a useful tool for confirmation of the prenatal diagnosis of mosaicism. Genetic analysis of multiple tissues is required postnatally.

Objective To investigate the action mechanism of IL-35 gene transfection ameliorating cardiac allograft rejection and prolonging allograft survival.Methods pEBI3-L-p35-Fc plasmid was amplified by polymerase chain reaction.In vitro plasmid DNA pEBI3-L-p35-Fc or pSec-L-Fc was,respectively,transfected into HEK293 cells using Lipofectamine 3000.At 48 and 72 h after transfection,IL-35 concentration in culture supernatant of transfected HEK293 cells was detected by ELISA.Balb/c and C57BL/6 splenocytes treated with mitomycin (MMC) served as the stimulators,those not treated with MMC as responders,and they were subjected to one-way mixed lymphocyte culture (MLC).In the presence or absence of IL-35,the percentage of CD4+ CD25+ Tregs was detected by flow cytometry.Abdominal heterotopic heart transplantation model was established by using inbred male Balb/c mice as donors and C57BL/6 as recipients respectively.In experimental group,recipients were intravenously administrated with IL-35 plasmid (50μg) on the day 1 to day 3 post-transplantation.The control mice were treated with normal saline.The IL-35 expression in the blood,CD4+ CD25+ Tregs proportion in the blood and spleen,and the survival and the histopathologic changes of the cardiac grafts were also observed.Results In vitro the transfected HEK293 cells expressed IL-35.IL-35 enhanced the proliferation of CD4+ CD25+ Tregs of MLC in vitro.The median survival time of the cardiac grafts in experimental group (16 days) was significantly longer than in control group (7 days) (P＜0.01).As compared with control group,CD4+ CD25+ Tregs proportion was significantly increased (P＜0.01),CD8+ T cells proportion was decreased (P＜0.01) and the proliferation of lymphocytes and monocytes infiltration was inhibited in the experimental group.Conclusion IL-35 could alleviate cardiac allograft rejection and prolong cardiac allograft survival via the induction of proliferation and differentiation of CD4+ CD25+ Tregs and inhibition of proliferation of CD8 + effector T cells.

Objective To investigate the expression of triggering receptor expressed on myeloid cells receptor-1 (TREM-1) in plasma and bronchoalveolar lavage fluid (BALF) and its correlation with Galactomannan,IFN,IL-6 and IL-10 in Aspergillus infected mice.Methods Cyclophosphamide (CTX) was intraperitoneally injected and fumigatus spore suspension was inhaled by nose to establish the immunocompromised invasive pulmonary aspergillosis (IPA) mouse model.Healthy controls,immunocompromised only and IPA only groups were also established.Each group had 6 mice.After inoculation,mice were sacrificed.Lung tissue specimens,BALF,and plasma samples were collected.Plasma and BALF soluble TREM-1 (sTREM-1),Galactomannan,IFNγ,IL-6,and IL-10 were detected by ELISA.Results Positive Aspergillus fumigatus was found by tissue culture in the lung.Infiltration of inflammatory cells,blood congestion and interstitial lung tissue injury were observed in histological sections of both IPA and immunocompromised IPA mice.Compared to IPA group [(453.78 ± 74.18) ng/L,P ＜ 0.001;(10.21±1.46) ng/L,P＜0.001] and control group [(245.16 ±65.85) ng/L,P＜0.001;(6.60 ± 3.74) ng/L,P ＜ 0.001],the plasma and BALF sTREM-1 significantly increased in immunocompromised IPA group [(1 537.64 ± 359.52) ng/L;(20.12-± 2.72) ng/L].Compared to control group,both the BALF sTREM-1 in IPA group (P =0.041) and the plasma and BALF Galactomannan,IFNγ,IL-6,and IL-10 levels in IPA and immunocompromised IPA groups were significantly higher (P ＜0.01).Pearson correlation analysis showed that plasma and BALF sTREM-1 were significantly correlated with Galactomannan (r =0.83,P ＜ 0.001;r =0.82,P ＜ 0.001),IFNγ (r =0.79,P＜0.001;r=0.61,P＜0.01),IL-6 (r=0.81,P＜0.001;r=0.66,P＜0.01),andIL-10 (r=0.70,P =0.001;r =0.54,P =0.02).Conclusions Plasma and BALF sTREM-1 appears highly expressed in Aspergillus infected mice.sTREM-1 in mice plasma and BALF is closely correlated with Galactomannan,IFNγ,IL-6,and IL-10 levels,which suggests that sTREM-1 has great diagnostic value during invasive fungal infection.

Objective To explore the research hotspots of critical care nursing in recent 5 years. Methods Based on PubMed, articles published during the last 5 years were retrieved using the MeSH termCritical Care Nursing. Bicomb 2.0 and SPSS 20.0 software were used to do word frequency analysis and co-word cluster analysis. Results A total of 826 articles were retrieved. With the top 42 keywords as high frequency keywords, 6 hotspots were extracted, including the allocation of critical care nurse staffing, attitudes and psychological experiences of people related to critical care nursing, competency of critical care nurses, evidence-based nursing practice, new techniques and methods of critical care nursing, and communication among critical care nurses, patients as well as patients′relatives. Conclusions Using co-word analysis, this study analyzed the hotspots of critical care nursing in recent 5 years. The results not only help researchers and clinical practitioners understand the current status of critical care nursing research, but also provide references for new developing studies and clinical practices.

The financial crisis has made the graduate of grade 2009 career have undergone profound changes in mentality,How to strengthen the ideological and political education of graduates and career guidance is a new task we have to face.In this paper,through a questionnaire survey,students’collective conversation,individual visits,the case summary,we understand the mental state of college graduates in 2009 session in Shenyan,and have analyzed it to provide a basis for career guidance for the targeted colleges and universities under the new circumstances.

Objective: To observe the clinical efficacy of long-snake moxibustion plus Western medicine for chronic heart failure due to heart-kidney yang deficiency.Methods: A total of 80 patients were randomized into a control group and an observation group, with 40 cases in each group. The control group was treated with routine Western medicine, and the observation group was treated with long-snake moxibustion therapy on the basis of routine Western medicine. Left ventricular ejection fraction (LVEF), stroke volume (SV) and left ventricular end-diastolic diameter (LVEDd) were measured before and after treatment, and the heart function was assessed. The serum brain natriuretic peptide (BNP) level was measured, the 6 min walking distance was recorded, and the Minnesota living with heart failure questionnaire (MLHFQ) was assessed and scored. The serum galectin-3 (Gal-3) and soluble growth stimulation expressed gene 2 (sST2) protein levels were measured. Clinical efficacy was evaluated after treatment. Results: The total effective rate of the observation group was higher than that of the control group, and the difference between the two groups was statistically significant (P<0.05). After treatment, LVEF and LVEDd in both groups were significantly improved (all P<0.05), and LVEF, SV and LVEDd in the observation group were superior to those in the control group (all P<0.05). The heart function classification in both groups were improved after treatment (both P<0.05), and the heart function classification in the observation group was superior to that in the control group (P<0.05). The serum BNP level in both groups decreased (both P<0.05), and the serum BNP level in the observation group was lower than that in the control group (P<0.05). The 6 min walking distance and MLHFQ scores in both groups were improved (all P<0.05), and the 6 min walking distance and MLHFQ score in the observation group were superior to those in the control group (both P<0.05). The serum Gal-3 and sST2 levels in the observation group decreased significantly (both P<0.05), and all lower than those in the control group (both P<0.05). Conclusion: Long-snake moxibustion plus Western medicine has significant efficacy in treating chronic heart failure due to heart-kidney yang deficiency; it can improve heart function, reduce the serum BNP level, and improve the exercise endurance and quality of life in the patients. This may be related to the down-regulation of serum Gal-3 and sST2 levels.