Objective:To identify the function of the 5′-flank upstream regulation region of human CD226 gene.Methods:The upstream regulation region of CD226 was cloned by PCR and ligated into pGL3 vector. Then the vector was transfected into Jurkat cell and luciferase activity was detected after 48 h culture.Results:CD226 gene may have two promoters, P1 and P2,which were located at the region of -843--319 bp and +1-+181 bp respectively, and PMA can up-regulate P1 while down-regulate P2. Both P1 and P2 can be up-regulated by A23187, especially P2.Conclusion:CD226 gene may have two promoters, and PMA and A23187 can regulate CD226 promoter activity in the similar pattern of protein level.

Objective To evaluate the clinical efficacy of dexmedetomidine given intranasally for day surgery recovery period.Methods 60 pediatric patients who were treated for elective day surgery were selected as study objects,and they were randomly divided into control group and observation group according to the digital table,30 cases in each group.During operation, after induction of 8% sevoflurane inhalation at 8 L/min,3% of sevoflurane was maintained at 1 L/min.After intubation,0.5 g dexmedetomidine was given intranasally in the observation group, 0.9% NaCl saline was given to the control group.Mean arterial pressure (MAP),heart rate (HR),oxygen saturation (SpO2) were observed at different time points.The recovery time, number of cases of agitation, Aono agitation scale, children anesthesia delirium PAED score and the incidence of adverse reactions were observed in the two groups.Results As the time passed by, the MAP,HR,SpO2 in the two groups stabilized, except HR of the observation group at T0 time was higher than the control group[(92.92±3.22)beats/min vs.(89.32±3.22) beats/min], the difference was statistically significant(t=2.34,P<0.05).The children of the observation group recovered more slowly than the control group[(20.55±0.97)min vs.(18.62±1.03)min,t=2.17,P<0.05)],the number of cases of agitation, Aono restless score, anesthesia delirium PAED scores were lower than children in the control group (t=5.38,3.72,2.56,all P>0.05).The overall incidence of adverse reactions was similar in the two groups, there was no statistically significant difference (13.33% vs.10.00%,x2=0.12,P>0.05).Conclusion Dexmedetomidine given intranasally for the children underwent day surgery can significantly improve the quality of children awakeness, reduce the incidence of postoperative delirium, and without significant side effects.

BACKGROUND: Host endothelial cells are important targets of alloreactive donor cytotoxic T lymphocytes in graft-versus-host disease (GVHD). And the expression of tissue factor in vascular endothelial cells is up-regulated post-injury and activation. So tissue factor expression in vascular endothelial cells may play a pivotal role in the pathogenesis of GVHD.OBJECTIVE: To investigate the molecular mechanism of allogeneic T cell-induced tissue factor (TF) expression in vascular endothelial cells.DESIGN, TIME AND SETTING: The cytology observation was performed at immunological laboratory of ongji Medical College, Huazhong University of Science and Technology from March to October 2008.MATERIALS: Human primary umbilical vein endothelial cells (HUVECs) were purchased from Science cell laboratory (CA, USA).METHODS: Allogeneic CD4~+ and CD8~+T cells were isolated from peripheral blood mononuclear cells (PBMC) by positive selection using magnetic beads coated with anti-CD4 or anti-CD8 antibody. After thorough washing with PBS, 1×10~6 allogeneic CD4~+T cells or CD8~+T cells were added to HUVECs (ratio 10:1). The cells were grown at 37 ℃ in a humidified atmosphere containing c-jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated protein kinase (ERK). HUVECs were pretreated with TF antibody 1 hour before co-culturing; the untreated HUVECs were used as controls. MAIN OUTCOME MEASURES: Allogeneic T cells-induced TF expression in vascular endothelial cells was determined by real-time PCR and flow cytometry. Western blot was used to examine allogeneic T cells-induced phosphorylation of MAPK in HUVEC.RESULTS: After co-culturing with allogeneic CD4~+T cells or CD8~+T cells for 6, 12, and 24 hours, the expression rates of TF on HUVEC membrane were obvious increased than that of the control group (P < 0.05), the expression levels of TF mRNA in HUVEC were also significantly elevated (P < 0.05). Allogeneic CD4~+T cells-induced expression rates of TF were significantly higher than allogeneic CD8~+T cells-induced expression rates at 12 and 24 hours after culture. The expression of p38MAPK and JNK was enhanced after culture with allogeneic T cells, yet the ERK expression had no changes. The p38MAPK or JNK can notably down regulated TF expression, which could block the TF expression when worked together. CONCLUSION: Allogeneic T cells induced TF expression in HUVECs via activation of p38MAPK and JNK.

Objective To explore the expression of protease-aetivated receptor (PAR)-1, 2 in a routine model of acute graft-versus-host disease (aGVHD). Methods A routine model of aGVHD after aUogeneic hematopoietic stem cell transplantation (allo-HSCT) was established, and the syngeneic HSCT mice were used as the controls. Quantitative real-time PCR, Western blot and immunohistoehemistry were done to detect the gene and protein expression of PAR-1, 2 in multiple organs of allo-HSCT mice and the controls. Results Allo-HSCT mice showed classical symptoms and histological changes of aGVHD. PAR-1 mRNA expression was significantly increased in the skin,liver, small intestine of allo-HSCT mice (skin: 0. 039 ± 0. 013 vs. controls: 0. 008 ± 0. 002,P<0. 01 liver: 0. 165 ± 0. 084, vs. controls: 0. 017 ± 0. 006, P<0. 01 ; small intestine: 0. 215 ± 0. 109 vs.controls: 0. 016±0. 009, P<0. 01), but not in the stomach, lung and kidney of allo-HSCT mice (P >0. 05). PAR-2 mRNA expression in the liver and small intestine of allo-HSCT mice was significantly elevated (liver: 0. 010 ± 0. 003 vs. controls: 0. 003 ± 0. 002, P<0. 01 ; small intestine:0. 006 ± 0. 002 vs. controls: 0. 003± 0. 002,P<0. 05), but not in the other organs (P>0. 05). The protein expression of PAR-1, 2 was in accordance with the mRNA expression. Immunohistochemistry revealed the PAR-1, 2 expression was increased in the epithelial eeUs and vascular endothelial cells of target organs of aGVHD. Conclusion Inereased expression of PAR-1, 2 in the target organs of aGVHD suggests PAR-1, 2 may contribute to the pathogenesis of aGVHD after allo-HSCT.

@@

Objective To evaluate the value of the tests of mean corpuscular volume (MCV ) ,mean corpuscular hemoglobin (MCH) in the screening of thalassemia .Methods 603 cases were performed MCV and MCH tests and gene detection of thalasse-mia .The results of gene detection were used as reference standard .The sensitivity ,specificity ,accuracy ,positive predictive value and negative predictive value of single or combined results of MCV ,MCH tests were calculated .Results The sensitivity and specificity of single MCV test were 91 .2% and 76 .1% ,single MCH test were 91 .2% and 70 .7% .The sensitivity and specificity of parallel combined tests of MCV and MCH were 94 .8% and 61 .4% ,serial combined tests of MCV and MCH were 88 .4% and 75 .6% .Con-clusion The sensitivity of single MCV or MCH test in the screening of thalassemia is high .Parallel combined tests of MCV and MCH can improve the sensitivity ,and will be widely used in the in the screening of thalassemia .

Objective To investigate the value of the interhospital transportation and the advantage of the cooperative network to the critically ill children.Methods The clinical data of 232 critically ill chil-dren transported from other hospitals in long distances to PICU of Fudan University Affiliated Children Hos-pital cooperated with Shanghai 120 interprovincial transport Department,between Oct 2010 and Oct 2013, were analyzed retrospectively.Results At otal of 232 critically ill childrne were transported from 31 hospi-tals of Eastern China region including five provinces and the city of Shanghai.Among thse e critically ill chil-dren 141 casse were male and 91 cases were femla e,with age from 29 days ot 13 years( median age was 12 months) and weight from 2.5 to 66 kg ( median weight was 10 kg ) .The average pediatric clinical illness score was 80.4 ±7.7,155 cases(66.8%) were single organ dysfunction,55 cases(23.7%) were multiple organs dysfunction,105 cases(45.3%) were mechanical ventilation required fort ransportait on.Referral radi-us was 50-1 000 km(median).Among these patients,87 cases(37.3%) weret ransported over 200 km radius and73 cases(36%) were transop rted 101-200 km,62 cases(26.7%) were transported 100 km or less.D ur-ing the transportation,0 case died,3 cases(1.3%) received cardiopulmonary resuscitation treatment,2 cases (0.9%) received electrical conversion treatment,2 cases(0.9%) were replaced endotracheal intuab tion,the success rate of transportation was 100%.The top 4 disae ses were respiratro y id seases(90 cases,38.8%), neural diseases(43 cases,18.5%) ,cardiovascular diseases(36 cases,15.5%) and acute pediatric trauma(28 cases,12.1%) .Al l 232 cir tically ill children were admitted to our PICU for further treatment through the Green Channel.In the end, 178 cases ( 76.7%) discharged, 24 cases ( 10.3%) gave up and 32 cases (13.8%) died.Among thse e 232 rc itically ill children,30 cases(12.9%) received special organ replacement treatment,4 casse ( 1.7%) of whom were treated with extracorporeal membrane oxyg enation, 26 cases (11.1%) received blood purification therapy.Conclusion Establishing the system of transportation between PICU of hospitals will be propitious to treat the critically ill children energetically and effectively.It is worth pro-moting to master the transportation indication of children with critical illness,which is safe and reliable.

Objective To investigate the effect of transforming growth factor-?1 (TGF-?1) on the activity of connective tissue growth factor(CTGF) gene promoter in human renal proximal tubular epithelial cell line HK-2. Methods The regulation fragment of 5' flanking region of human CTGF gene was linked to pGL3-Basic vector. The recombinant plasmid pCTGF-luc was transient transfected to HK-2 cells. The activity of CTGF promoter after treatment of TGF-?1 and mitogen-activated prontein kinases (MAPK) pathway inhibitors was assayed by means of luciferase reporter gene assay system. Results TGF-?1-induced increase of CTGF promoter activity was concentration-dependent, with a plateau at 5 ng/ml by 1.82-fold vs control (P

Objective To investigate the expression of vascular endothelial factor C(VEGF-C) and E-cadherin in gastric cancer and explore the relationship between expression of VEGF-C and E-cadherin and the pathogenesis of gastric cancer.Methods Real-time quantitative reverse ranscriptase-polymerase chain rection was applied on 40 cases of gastric cancer and corresponding adjacent noncancerous tissues,in order to detect mRNA expression of VEGF-C and E-cadherin gene.VEGF-C and E-cadherin protein expression in gastric cancer and adjacent normal gastric mucosa were detected by immunohistochemistry.Statistical analysis was carried out to analyze the correlation among VEGF-C,E-cadherin and protein expression with various clinical parameters in these gastric cancer patients.Results The expression of VEGF-C protein in 40 cases of gastric cancer's cancer tissues was 0.461±0.012,significantly higher than that in the adjacent normal tissues(0.036+0.023;t=1.101,P＜0.05),but E-cadherin expression was significantly lower than that of the adjacent normal tissues (0.079±0.002 vs.0.321±0.005;t=1.844,P＜0.05).There was correlation between VEGF-C mRNA expression with histological grading,TNM staging,lymph node and distant metastasis (t=-1.621,-1.474,-2.378,-1.966,P＜0.05).There was correlation between E-cadherin mRNA expression with tumor size,histological grading,TNM stage,there was a significant difference (t=1.875,1.673,1.544,P＜0.05).VEGF-C and E-cadherin protein expression was negatively correlated(r=-0.688,P＜0.05).Conclusion Up-regulated of VEGF-C gene and decreased expression of E-cadherin may play an important role in the carcinogenesis of gastric cancer

Fifteen patients with chronic renal failure(CRF)underwent laparoscopic cholecystectomy(LC)at carbon dioxide(CO2)pneumoperitoneum pressure of 10-12 mm Hg(Group A,n=9)or 13-15 mm Hg(Group B,n=6).Renal function and urinary volume(UV)of Group A showed no remarkable change following the operation.But in Group B,the levels of blood urine nitrogen(BUN)and serum creatine(Scr)were increased significantly,and creatinine clearance rate(Ccr)and UV were remarkably decreased(P＜0.05).These variants gradually retumed tO the preoperative levels after 1 week.The analysis showed that laparoscopic choleeystectomy at CO2 pneumoperitoneum pressure of 10-12 mm Hg in CRF patients might be safe.Higher CO2 pneumoperitoneum pressure could result in reversible renal Email:yuanhuizhong2000@yahoo.com.cnfunction change.