Objective:To clone and functionally analyze the promoter region of an intestinal specific expressed gene,Musashi-1.Methods:The 5' flanking region of MSI-1 gene was cloned from C57BL/6J mouse genomic DNA using PCR-mediated recombination.Expression of MSI-1 mRNA was determined in Colon26 and B16 cell lines using Northern Blotting.Various 5′-deletion recombination plasmids were constructed and transfected transiently into the Colon26 cell line.Luciferase reporter assay was performed to determine the relative transcriptional activities of various 5′-deltion fragments.Results:MSI-1 mRNA was expressed in both Colon26 and B16 cell lines,but much higher in Colon26 cell line.The transcriptional activity of DNA fragment from 73bp downstream to 4 939bp upstream the(MSI-1) gene transcriptional start site was 29.9 fold of the pGL3-basic empty vector.Conclusion:(pSMI+73~)-4 939 has the transcriptional activity and can be regarded as the promoter of MSI-1 gene.A cis-acting element lies between 4 011bp to 4 939 bp upstream the transcriptional start site of(MSI-1) gene,which may be responsible for the tissue specific expression of MSI-1 gene.The cloning of MSI-1 gene promoter is a precondition for the isolation and purification of intestinal stem cells using this promoter.

Altering the natural history of disease has been the goal of medical therapy for Crohn's Disease (CD) in recent years. The conventional approach has been thestep-up therapy. With the introduction of biological agents (such as infliximab) , there is growing evidence that the top-down approach is justified in a subgroup of high-risk patients. The new algorithms is not only associated with superior clinical remission rates, but also may have the potential to alter the natural history of the disease, including inducing mucosal healing, reducing the rate of complications, surgeries and mortality. Further study will have to determine which subgroup of patients will benefit most from this early aggressive biological therapy approach.

With the increasing understanding of the pathogenesis of Crohn's disease and the advent of biological therapy,the therapeutic strategies of Crohn's disease have changed a lot in recent years.In this review,we concerned the progress in the management of active disease,maintenance of medically induced remission,and surgical options of Crohn's disease.The new concept of "top-down" approach to the treatment of Crohn's disease evolved recently was also discussed.

Abdominal compartment syndrome(ACS)is an important factor contributing to the multiple organ dysfunction syndrome which is commonly seen in patients with severe acute pancreatitis(SAP).As a life-saving procedure,decompressive laparotomy is widely applied to patients with SAP complicated with ACS,especially to patients with edema of the visceral tissues caused by massive fluid resuscitation at the early stage of the disease.However,decompressive laparotomy should be adopted with caution since it is associated with enteroatmospherie fistula,intraabdominal infection,postoperative ileus,third space losses,hypothermia and hemia.Therefore,decompressive laparotomy should only be considered after conservative management had failed.The indications for decompressive laparotomy are as follows:(1)intraabdominal pressure＞25 nnn Hg;(2)adequate ventilation of the patient is difficult;(3)pereutaneous drainage of ascites is not helpful.Timely temporal abdominal closure is helpful in preventing complications.Infected peripanereatie necrosis is the indication for peripancreatic exploration or necroseetomy.A thorough knowledge of decompressive laparotomy is essential for individualized management of patients with SAP complicated with ACS.

Altering the natural history of disease has been the goal of medical therapy for Crohn's Disease(CD) in recent years.The conventional approach has been the "step-up" therapy.With the introduction of biological agents (such as infliximab),there is growing evidence that the "top-down" approach is justified in a subgroup of high-risk patients.The new algorithms is not only associated with superior clinical remission rates,but also may have the potential to alter the natural history of the disease,including inducing mucosal healing,reducing the rate of complications,surgeries and mortality.Further study will have to determine which subgroup of patients will benefit most from this early aggressive biological therapy approach.

In order to explore whether gene CHFR was inactivated by methylation in leukemia cells, the expression of CHFR was examined before and after treatment with demethylation agent in Molt-4, Jurkat and U937 leukemia cell lines by means of RT-PCR. The methylation of promoter in Molt-4, Jurkat and U937 cells as well as 41 acute leukemia patients was analyzed by MS-PCR. The results showed that methylation of CHFR promoter was inactivated and could be reversed by treatment with a demethylating agent in Molt-4, Jurkat and U937. CHFR promoter methylation was detected in 39% of acute leukemia patients. There was no difference in incidence of CHFR promoter methylation between acute myelocytic leukemia and acute lymphocytic leukemia. In conclusion, CHFR is frequently inactivated in acute leukemia and is a good candidate for the leukemia supper gene. By affecting mitotic checkpoint function, CHFR inactivation likely plays a key role in tumorigenesis in acute leukemia. Moreover, the methylation of gene CHFR appears to be a good index with which to predict the sensitivity of acute leukemia to microtubule inhibitors.
Cell Cycle Proteins/*genetics ; DNA Methylation ; DNA, Neoplasm ; Epigenesis, Genetic ; Leukemia, Myeloid, Acute/*genetics ; Neoplasm Proteins/*genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/*genetics ; Promoter Regions (Genetics)/*genetics ; Tumor Cells, Cultured

Objective To investigate the clinicial significance of continuous glucose monitoring(CGM)of patients with severe traumatic brain injury(sTBI). Methods By glucose monitoring method,80 patients with sTBI〔Glasgow coma score(GCS)3-8〕in Department of Critical Care Medicine of Qingyuan People's Hospital in Guangdong Province from January 2012 to December 2012 were divided into two groups:41 patients in CGM group and 39 in regular glucose monitoring(RGM)group. The continuous glucose monitoring system(CGMS)was applied to monitor glucose level in the CGM group,and the finger blood was taken by portable blood glucose meter in the RGM group. The two groups were treated with insulin on the basis of glucose level,respectively. The relationships between the condition of glycemic excursions and the acute physiology and chronic health evaluationⅡ(APACHEⅡ)score or prognosis and between the incidence of hypoglycemia and prognosis were seen in the two groups. Results The close linear correlations between APACHEⅡ score and glycemic excursion in two groups,i.e. mean amplitude of glycemic excursions(MAGE)and coefficient of variation of glucose(GluCV),were documented(both P<0.05). The MAGE of the especially severe patients(GCS 3-5)was obviously higher than that of severe ones(GCS 6-8),and with the increase of APACHEⅡ score,the MAGE of patients was gradually elevated,the difference being statistically significant(both P<0.05). The incidence of hypoglycemia(7.32%vs. 23.08%)and fatality rate of 30 days(12.20%vs. 30.77%)in CGM group were lower than those of RGM group(both P<0.05). The MAGE and fatality rate of 30 days were positively correlated in CGM group(r=0.597,P=0.007),and the GLuCV and fatality rate of 30 days were positively correlated in RGM group(r=0.622,P=0.019). Conclusion CGM is beneficial to timely observe condition of glycemic excursions in sTBI patients and avoid occurrence of hypoglycemia or hyperglycemia,guiding the treatment of insulin and improving patients' prognosis.

Objective To explore the diagnosis,treatment,risk factors and prognosis factors of postpancreaticoduodenectomy hemorrhage (PPH).Methods The retrospective case-control study was adopted.The clinical data of 703 patients who underwent pancreatoduodenectomy at Hospital 401 of the People's Liberation Army from January 2008 to July 2013 were collected.Standard pancreatoduodenectomy was carried out for the malignant tumors of the head of pancreas or ampulla,pylorus-preserving pancreatoduodenectomy was operated for the benign tumor or the duodenal papilla tumor.The corresponding treatment was adopted for PPH.The observation indicators included:(1) the surgical situation (surgical method,operation time and the volume of intraoperative blood loss),(2) diagnosis of PPH,(3) treatment of PPH,(4) univariate and multivariate analyses for the risk factors affecting the occurrence of PPH,(5) univariate and multivariate analyses for the risk factors affecting prognosis of PPH patients.The measurement data with normal distribution were represented as x ± s.The measurement data with skewed distribution were represented as M (range).The chi-square test or Fisher exact probability was used for univariate analysis.Logistic regression model was used for multivariate analysis.Results (1) The surgical situation:among 703 patients,409 patients underwent standard pancreatoduodenectomy and 294 underwent pylorus-preserving pancreatoduodenectomy,including 1 combined with right hemihepatectomy,27 with portal vein reconstruction and 2 with hepatic artery reconstruction.Pancreaticojejunostomy was applied to 658 patients using mucosa anastomosis of the pancreatic duct to jejunum and 45 patients using invagination anastomosis.Supporting tube was routinely deposed in the pancreatic duct,598 patients had internal drainage and 105 patients had external drainage.The end-to-side anastomosis between common bile duct and jejunum was used for choledochojejunostomy.The 409 patients received the gastrojejunostomy using side-to-side anastomosis of gastric part and jejunum and 294 patients using end-to-side anastomosis of duodenum and jejunum.Operation time and volume of intraoperative blood loss were (324 ± 54) minutes and (428 ± 118) mL.(2) The diagnosis of PPH:among 703 patients after pancreatoduodenectomy,62 patients had PPH,the hemorrhage reasons of 38 patients had been identified,and the hemorrhage reasons of 24 patients had not been identified (A level in 5 patients,B level in 17 patients,C level in 2 patients).① The site of hemorrhage:the hemorrhage outside the cavity were detect in 27 patients,the hemorrhage inside the cavity in 28 patients,and the hemorrhage from both outside and inside part of the cavity in 7 patients.② The time of hemorrhage:early-stage hemorrhage were detected in 5 patients and the delayed hemorrhage in 57 patients.③The volume of postoperative blood loss was (885 ± 253)mL,30 patients had mild hemorrhage and 32 patients had severe hemorrhage.④ The clinical classification of PPH:5,32 and 25 patients were detected in level A,B,C,and 19 patients combined with sentinel hemorrhage.(3) The treatment of PPH:①5 patients with PPH in A level were given clinical observation,blood volume supplement and other treatment,then the symptoms gradually turned better.② Among 32 patients with PPH in B level,15 patients became better after symptomatic and supportive treatments,6 patients received successful hemostasis after guglielmi detachable colis embolization,4 patients received successful hemostasis under gastroscopic hemostasis,7 patients received emergency exploratory laparotomy.Thirty-two patients were improved and then out of hospital after treatment,without occurrence of death.③ Among 25 patients with PPH in C level,4 patients received successful hemostasis after guglielmi detachable colis embolization,17 patients received hemostasis by emergency exploratory laparotomy,4 patients with undiscovered bleeding points received the treatment of fluid infusion,blood volume supplement and antacid.Among 25 patients after corresponding treatment,10 patients were improved and 15 patients were dead.(4) The result of univariate analysis showed that the combined hypertension,vascular resection and reconstruction,postoperative pancreatic leakage and postoperative intraabdominal infection were risk factors affecting the occurrence of PPH (x2 =4.950,5.300,7.568,5.505,P ＜ 0.05).The results of multivariate analysis showed that the combined pancreatic leakage and postoperative intraabdominal infection were independent risk factors affecting the occurrence of PPH [OR =2.761,2.216,95％ confidence interval (CI):1.389-5.489,1.198-4.101,P ＜ 0.05].(5) The risk factors affecting the prognosis of PPH patients:the results of univariate analysis showed that postoperative sentinel hemorrhage,postoperative pancreatic leakage,site,degree and level of hemorrhage were risk factors affecting the prognosis of PPH patients (x2 =8.022,4.448,11.853,18.551,28.285,P ＜ 0.05).The results of multivariate analysis showed that postoperative sentinel hemorrhage and site of hemorrhage (outside and inside part of the cavity) were independent risk factors affecting the prognosis of PPH patients (OR =5.550,0.233,95％ CI:1.595-19.314,0.086-0.635,P ＜ 0.05).Conclusions Pancreatic leakage and intraabdominal infection are independent risk factors after pancreatoduodenectomy.The treatment effect of the early-stage hemorrhage is better than that of the delayed hemorrhage,and angiographic embolization is the first choice of diagnosis and treatment for the delayed hemorrhage.Sentinel hemorrhage could result from aneurysm or continuous arterial hemorrhage of vascular erosion,it is the independent risk factor affecting the death of hemorrhage after pancreatoduodenectomy.

Objective To observe the effects of tetramethylpyrazine on intraoperative autotransfusion of blood quality , analysis of ligustrazine in autotransfusion of clinical value of blood quality control.Methods 68 patients undergoing elective surgery in our hospital from February 2015 to May 2016 were selected as the research object,and were randomly divided into control group and experimental group,each group of 34 cases.Two groups of patients in the intraoperative autologous blood transfusion and the control group according to conventional autologous transfusion in operation ,test group of patients with intravenous injection of ligustrazine injection in the recovery of blood before adding ligustrazine injection in the recovery process of blood.The two groups were collected before blood transfusion,observe the morphology of red blood cells, red blood cell fragments under the microscope and compared;the two groups were collected after reinfusion of blood two hours,one and five days in peripheral venous blood, the determination of superoxide dismutase (SOD), malondialdehyde (MDA), T lymphocyte subsets level of two groups were compared before and after one and five days. Results The erythrocyte deformability and erythrocyte debris of the two groups were higher than those before operation (P<0.05).The blood red blood cell deformability and the number of red blood cell debris in the blood transfusion group were significantly higher than those in the control group Significantly lower than the control group (P<0.05).The levels of SOD were significantly lower at two hours and one day after operation than those before operation, and the serum SOD was still lower than that before operation in the control group,The SOD in the experimental group was significantly higher at two hours, one and five days in the control group,MDA was lower than the control group(P<0.05).The CD3 +,CD4 +, CD4 +/CD8 +levels were significantly lower at two hours and one day after operation than those before operation (P<0.05).CD3 +,CD4 +and CD4 +/CD8 +in the control group were still lower than those in the control group at five days after operation(P<0.05),the levels of CD3 +, CD4 +, CD4 +/CD8 +were significantly higher in the experimental group than those in the control group at two hours,one and five days after treatmen (P <0.05).There was no significant difference between the two groups in the incidence of adverse reactions.Conclusion The addition of preoperative intravenous injection of ligustrazine,blood recovery process,can effectively protect the transfusion of red blood cell integrity,reduce the effect of transfusion of blood and blood antioxidant capacity in patients with T lymphocyte immune function ,to improve the quality of blood transfusion patients has important clinical value .

This study was designed to determine the impact of chrysoeriol on proliferation and cell cycle progression in the human multiple myeloma cell lines RPMI 8226 and KM3, and its related molecular mechanisms. Chryseoriol was identified by using the phosphorylated AKT-specific cytoblot high throughput assay. CCK-8 assay was employed to examine the growth inhibition rate and IC(50) (48 h) in peripheral blood mononuclear cells (PBMNCs), RPMI 8226 and KM3 cells treated with chrysoeriol at various concentrations. Cells were labeled with 5-6-carboxyfluorescein diacetate succinimidyl ester (CFSE), and the proliferation dynamics was detected by flow cytometry and analyzed with ModFit software. The cell cycles of RPMI 8226 and KM3 cells were measured by flow cytometry when the IC(50) concentration of chrysoeriol was adopted. The alterations in cell-cycle related proteins (Cyclin B1, Cyclin D1, p21) and proteins in PI3K-AKT-mTOR pathway were determined by Western blot analysis. The results showed the proliferation of multiple myeloma cells was significantly inhibited by chrysoeriol, resulting in cell cycle arrest in G(2)/M phase. Chrysoeriol could significantly reduce the expression of p-AKT (s473) and p-4eBP1 (t37/46) protein, meanwhile enhanced Cyclin B1 and p21 protein expression. Similar effects were not observed in PBMNCs from normal donors. It was concluded that chrysoeriol was a selective PI3K-AKT-mTOR pathway inhibitor. It restrained the proliferation of human multiple myeloma cells, but didn't affect proliferation of PBMNCs from normal donors. It might exhibit the cell cycle regulatory effect via the inhibition of PI3K-AKT-mTOR signal pathway.