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To investigate the expressions of Bcl-2 and Beclin-1 in pancreatic cancer and analyze the correlation between them.The pancreatic tissue samples were collected from each 6 cases of pancreatic cancer, pancreatic exocrine benign tumor, chronic pancreatitis and normal pancreas and marked as group A, group B, group C and group D, respectively. The mRNA expression levels of Bcl-2 and Beclin-1 were detected by real-time fluorescence quantitative PCR and the protein expression levels of Bcl-2 and Beclin-1 were detected through immunohistochemistry.The expression levels of Bcl-2 mRNA and protein, were the lowest in group D and the highest in group A (P0.05). The expression levels of Beclin-1 mRNA and protein in group A were significantly lower than those in group B and group D (P0.05). However, the expression levels of Beclin-1 between group A and group C were not significantly different (P0.05). The correlation coefficient between Bcl-2 and Beclin-1 protein expression in pancreatic cancer is--0.827 (P = 0. 042).Compared with normal pancreatic tissue, pancreatic cancer had Bcl-2 expression upregulated and Beclin-1 expression downregulated. The increased anti-apoptotic effect of Bcl-2 and the decreased autophagic effect of Beclin-1 may collaboratively contribute to the occurrence of pancreatic cancer.

Background/aims The timing for the management of gallstones pancreatitis remains a contentious issue. Various scholars have their own achievement in in regards to this issue. Methodology We reviewed our hospital charts from Jan 2007 to December 2010 and made a comparative study about early and delayed LC for mild to moderate gallstone pancreatitis in 80 patients. Results Successful management was obtained in all patients. Out of 80 patients, 54 had underwent for early LC within 48 hours and 26 delayed LC (6-8 weeks). Conclusions Our study reveals that early cholecystectomy has nice outcomes in terms of shorter hospital stay and expenses. Proper consultation should be taken from radiological colleague if CBD dilations are >6 mm and contraction of gallbladder appears on imaging modalities. Comorbid conditions, past history of cholecystitis cannot be avoided for proper surgical outcomes. Postoperative complications can be deterred by early LC for mild gallstone pancreatitis. However, large volume studies are essential from different places to answer the debated topic of which management protocol is justifiable for the management of mild to moderate gall stone pancreatitis.

The purpose of this study was to establish the phospho-specific flow cytometry (phospho-flow) to detect the phosphorylated signaling proteins of leukemia cells and to evaluate its useful value in leukemia study. The bone marrow of leukemia children was collected, and treated by phospho-flow of extracted mononuclear cells (MNC) and phospho-flow of directly fixed bone marrow (BM) respectively. In phospho-flow of extracted MNC, the MNC extracted from BM were fixed and permeabilized, then were cultured with P-AKT and P-ERK1/2, finally were analyzed by flow cytometry. In phospho-flow of directly fixed BM, the BM was treated with fixation/lysis buffer and 90% methanol, then were incubated with the surface CD antibody, P-AKT and P-ERK1/2, finally the treated BM cells were analyzed by flow cytometry. The results showed that the positive rates of P-AKT and P-ERK1/2 in MNC treated by phospho-flow of extracted MNC of 26 leukemia children were 46.2% and 30.8% respectively, while the positive rates of P-AKT and P-ERK1/2 in BM treated by phospho-flow of directly fixed BM were 50.0% and 38.5% respectively. The comparison of positive rates of P-AKT and P-ERK1/2 between the 2 treatment protocol showed no difference (p0.05). It is concluded that the phospho-flow of directly fixed BM established by our laboratory can be used to analyze the signaling proteins of leukemia cells.

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To study the clinical significance of Toll-like receptor 4 (TLR4) in children with severe sepsis.A prospective control study was performed. All cases were enrolled from pediatric department of the First Affiliated Hospital of Guangzhou Medical College, and they were divided into severe sepsis group (14 patients) who were diagnosed to have severe sepsis or septic shock in intensive care unit (ICU), pneumonia group (10 cases) with diagnosis of bronchial pneumonia, and healthy control group (10 healthy children). Venous blood samples of 2 ml were collected at admission, the level of TLR4 was detected by flow cytometry .At the same time, the changes in serum interleukin (IL-6, IL-10 ) and tumor necrosis factor-α (TNF-α) levels were determined by enzyme linked immunoadsorbent assay (ELISA).In severe sepsis group, the contents of TLR4 [(71.56±15.32)%], IL-6 [(1.98±1.55) ng/L], IL-10 [(88.20±61.23) ng/L] and TNF-α [(104.08±85.36) ng/L] were significantly higher than those in pneumonia group [(50.07±26.36)%, (0.93±0.16) ng/L, (41.42±7.02) ng/L, (48.96±6.40) ng/L] and healthy control group [(39.43±17.43)%, (0.94±0.43) ng/L,(43.73±22.68) ng/L, ( 49.94± 18.47) ng/L, all P0.05). But there was no significant difference in the contents of TLR4, IL-6, IL-10 and TNF-α between pneumonia group and healthy control group (all P0.05).This study suggests that TLR4 might be critically involved in the development of sepsis, and changes in TLR4 expression are parallel with levels of proinflammatory cytokines, including IL-6, TNF-α, and IL-10. The combination of TLR4 and proinflammatory cytokines would serve as the predictive parameters in early diagnosis and severity evaluation of sepsis in children.

To explore the molecular mechanism of interleukin cytokines IL-6, IL-17, IL-23, IL-23R in the onset of acute pancreatitis in rats pancreatitis models.Thirty six SD rats were randomized into 3 groups: control group, model group, and intervention group (n = 12). The rats in model group and intervention group were induced by intraperitoneal injection of 1-arginine, and those in intervention group were treated by tail intravenous administration of drugs at the same time. Then the rats were sacrified at 3rd, 6th, and 12th h. after the modeling. The levels of IL-6 ,IL-17, IL-23, IL-23R in blood, pancreas, lung and kidney were checked by ELISA.The IL-6 levels of serum, pancreas, lung and kidney in AP Model group were obviously higher than those in the control group and intervention group. This trend increased with time. Similarly, the levels of IL-17 and IL-23 in AP group were obviously higher than those in the control group and intervention group. Compared with the control and intervention group, AP group showed higher IL-23R levels in serum, pancreas, but lower IL-23R levels in lung and kidney.IL-23 is a mediator involved in the formation of IL-17 and IL-6, they all can promote acute pancreatitis.

To investigate the effects of NKG2D mAb on the survival of allogeneic transplanted islets nonobese diabetic (NOD) mice, and to find if CD154 mAb has synergistic effects.Spontaneous diabetic NOD mice transplanted with allogeneic islets of BALB/c mice were divided into 4 groups. Group A was control group, Group B were treated with anti-NKG2D monoclonal antibody (mAb), Group C were treated with CD154 mAb (MR1), Group D were treated with NKG2D mAb and MR1. Glucose levels were monitored at regular intervals through caudal vein, and islet function was evaluated by glycemia. Histological study was performed at graft rejection or at day 120. Spleen cell suspension was prepared for mixed lymphocyte cultivation. The kidneys hosting the islet graft were prepared with HE staining and immuno-histochemistry staining of CD3, CD4 and CD8 was performed.MR1 therapy alone significantly prolonged the survival of islet grafts when compared to NKG2D mAb group and the control group: median graft survival was 41 days versus 8 days (P0.05) and 8 days (P0.05), respectively. Combination therapy with NKG2D mAb and MR1 prolonged islet grafts survival when compared to MR1 therapy alone: median graft survival was 51 days versus 41 days (P0.05).NKG2D mAb alone did not result in the prolongation of islet graft survival, whereas CD154 mAb increased graft survival. When both antibodies were administered, a synergistic effect was obtained, but did not provide permanent protection from diabetes recurrence.

To study the role of Th17 cells in the Immune rejection of islet transplantation, explore the feasibility of immune tolerance of islet transplantation induced by the combination applying of IL-23R antibody and Anti-CD154mAb.The in vitro experiments were divided into 5 groups: Blank control group, SD rat islet cells were cultured alone; A group, co-culture of rat pancreatic islet cells and lymphocytes, without IL-23R antibodies; B, C, D groups, co-culture of rat pancreatic islet cells and lymphocytes, respectively with IL-23R antibodies 0.1 microg/mL, 0.5 microg/mL, 1.0 microg/mL. Cells were harvested for Acridine orange (AO)/propidium iodide (PD) fluorescence staining, insulin and glucagon staining and glucose-stimulated insulin secretion test. The in vivo experiments (the purified islet to be transplanted under the left kidney capsule of the mice) were divided into four groups: Control group, BABL/c mice were transplanted with islets of SD rats with no treatments, IL-23R antibody (200 microg) treatment alone, anti-CD154mAb (200 microg) treatment alone and a combination of both. The blood glucose of the transplanted mice were monitored. The kidney of islet grafts were sliced for HE staining and insulin and glucagon immunohistochemical detection.Three days after mixed cultivation, the glucose stimulation index was 3.66 +/- 0.10 in blank control group, which was higher than that of other groups. Stimulation index of D group was 1.95 +/- 0.75, which was significantly higher than that of other groups. The functional graft survival of all experimental groups were significantly better than that of control group as demonstrated by immunohistochemical staining of insulin and glucagon, as well as in vitro and in vivo experiments. After three days of islet transplantation, the blood glucose of control group was higher than that of experimental groups, but no significant difference was observed among experimental groups.Th17 cells were involved in the islet transplant rejection. The expression of IL-17 could be considerably reduced through the block of the IL-23R, the effect of the block had a positive correlation in a dose-dependent manner. The combination of Anti-CD154 mAb and IL-23R antibody could prevent the acute rejection to some extent. However, there's no significant difference compared with the Anti-CD154mAb alone.

This study was purposed to investigate the changes of CD4(+) CD25(+) regulatory T cells and NK cells in peripheral blood of acute leukemia children at different stages, the function of immune system and the possible roles of the CD4(+) CD25(+) regulatory T cells as well as NK cells in leukemia immunity. The number and proportion of CD4(+) CD25(+) regulatory T cells and NK cells were detected by flow cytometry in the peripheral blood of 53 acute leukemia children, including 25 patients in new diagnosis and 28 patients in continuous complete remission (CCR), and were compared with that of 20 normal children. The results indicated that the mean proportion of CD4(+) CD25(+) CD127(+) in CD4(+) T cells of peripheral blood in newly diagnosed patients, patients with CCR and normal children were (9.55 +/- 2.41)%, (8.54 +/- 2.51)% and (6.25 +/- 0.85)% respectively, the mean proportions of CD4(+)CD25(+)CD127(+) in newly diagnosed patients and patients with CCR were higher than that in normal children, the mean proportion of CD4(+)CD25(+)CD127(+) in newly diagnosed patients were higher than that in patients with CCR (p0.05). At the same time, the NK cell count in patients with acute leukaemia decreased as compared with normal control, while after achieving CCR, the NK cell count in patients were also less than that in normal control (4.11 +/- 3.87% and 10.41 +/- 7.20% vs 14.06 +/- 5.95%, p0.05). It is concluded that the application of CD4(+), CD25(+) and CD127(+) to detect regulatory T cells is a simple, reproductive and accurate method, and the CD4(+) CD25(+) CD127(+) T cells can better reflect the proportion of CD4(+)CD25(+) regulatory T cells. The increase of regulatory T cells and decrease of NK cells in pediatric patients with acute leukemia indicate that the function of NK cells may be depressed. Treg T cells play a role in occurrence and development of leukemia, and are involved in down-regulating NK cell function.

To explore the value of the POSSUM scoring system in predicting postoperative morbidity and mortality of pancreatoduodenectomy (PD).Two hundreds and sixty-five consecutive PDs were performed between January 2005 and December 2007. POSSUM scores which relied on 12 physiologic and 6 operative variables were prospectively calculated for each case. Expected morbidity and mortality were estimated based on POSSUM scores and were compared with observed morbidity, which were diagnosed according to the Clavien complication scheme and domestic reference criteria respectively, and mortality.Physiologic scores of 265 cases ranged from 12 to 24,the mean was 15. Operative scores ranged from 14 to 24, the mean was 17. The overall POSSUM scores ranged from 0.24 to 0.88. Average expected morbidity was 43.8%, expected cases were 116. Observed morbidity rate was 39.6% (105/265). The expected and observed morbidities and cases had no significantly differences. All patients were classified to 1 of 4 strata based on their individual POSSUM scores and subsequent risk of morbidity. Predictive value was the highest when scores ranged from 0.4 to 0.8. POSSUM exhibited less predictive value for mortality, but if POSSUM was more than 0.5, it was useful for mortality predicting.POSSUM scoring system has high value for predicting the risk of morbidity in PD and can be helpful in guiding surgery and postoperative management decisions.

A new approach for the design of dichromatic white light-Emitting Diodes (LEDs) has been proposed by employing InAlGaN irregular multiple quantum well (IMQW) structures. The InAlGaN IMQW structures are assembled by two different type QWs emitting complementary wavelengths. The electronic and optical properties of the designed InAlGaN IMQWs have been analyzed in details by fully considering the effects of strain, well-coupling, valence band-mixing and quasi-bound states using a newly developed theoretical model from the k·p theory. The influence of material components, well width and well number of different type QWs, and barrier thickness on the spontaneous emission spectra of InAlGaN IMQWs was studied. The IMQW structure was obtained which can realize near white light emission by optimizing the structure parameters of IMQW structure for dichromatic white LEDs.

This study was aimed to investigate the conditions of culturing in vitro mesenchymal stem cells (MSCs) derived from bone marrow of children with acute leukemia and the biological characteristics of MSCs from leukemia children. The bone marrow MSCs of acute leukemia children were isolated by density gradient centrifugation combined with adherent segregating method and cultured in DMEM/F12. The morphology of Wright stained MSCs was observed under inverted microscope. Cell surface markers were analyzed with flow cytometry. The growth characteristic features of cultured MSCs was measured with MTT method. Induced adipogenic and osteogenic differentiation of MSCs in appropriate induction media was observed. The results indicated that BM-MSCs of acute leukemia children could be successfully cultured in vitro in appropriate conditions. At 24 hours of culture the MSCs began to adhere to wall, grew in colony and appeared in different shapes. As the culture lasted, the MSCs proliferated continuously and shaped in fusiform. After 2 - 3 weeks of culture, MSCs covered the bottom of culture flask. The analysis of growth feature showed that MSCs were in latency for 3 days, and then entered into growth period. After 8 days of culture the growth of MSCs showed to be in plateau stage. The shape of MSCs in 1st and 2nd generation showed to be heterogeneous but the 3rd generation to be homogeneous with long-fusiform. Cells were arranged in shape of whirlpool or radiation. The surface marker analysis showed that the MSCs were positive for CD105, CD29, CD13, but negative for CD34, CD45, CD14 and HLA-DR. The MSCs from leukemia children could be induced into adipocytes and osteocytes in appropriate conditions. It is concluded that (1) MSCs derived from children with acute leukemia can be successfully cultured and passaged in vitro; (2) MSCs from leukemia children not received chemotherapy are more successfully cultured in vitro than those received chemotherapy; (3) the common biological characteristics of MSCs from children with acute leukemia are same as the MSCs from healthy person.

Objectives In this study, we determined whether preoperative biliary drainage should be routinely performed in patients with jaundice. Methods The 342 patients undergoing pancreaticoduodenectomy between June 2004 and June 2008 were analyzed. Of these patients, 303 without biliary drainage were divided into 4 groups: (1) no jaundice, (2) mild jaundice, (3) moderate jaundice, and (4) severe jaundice. Multiple preoperative, intraoperative, and postoperative variables were examined. Postoperative complications were stratified by severity according to the modified Clavien classification. Results Patients with jaundice had a higher incidence in subsequent complications than those with no jaundice. The complications were stratified by severity. Compared with those in group 1, patients in groups 2, 3, and 4 had more complications just in grade 2 (15.6%, 23.1%, 28.3%, and 40.0%, respectively, P = 0.046), but not other more severe grades including 3a, 3b, 4a, 4b, and 5; all of the complications in this grade could be conservatively treated and cured without requiring surgical, endoscopic, or radiological intervention. The incidences of infection and overall complications were higher in patients with drainage than those without, but neither difference was statistically significant. Conclusions Preoperative drainage should not routinely be performed in patients with jaundice scheduled for pancreaticoduodenectomy, and immediate surgery is preferable.

To assess the value of perforin mRNA and granzyme B mRNA in lymphocytes as an early diagnostic indicator for acute rejection of pancreatic transplantation.Twenty-four hybrid landraces were divided into three groups (control group, transplantation group, transplantation plus immunosuppressant group) randomly. The landraces in the transplantation group were treated with pancreatoduodenal allotranplantation. The landraces in the transplantation plus immunosuppressant group underwent the same surgery and were intervened simultaneously with CsA, MMF, and Metrisone. Peripheral venous bloods were sampled via subclavic veins one day before the operations and on the 1st, 3rd, 5th and 7th day after the operations. The RNA was extracted from the blood samples. The mRNA of perforin and granzyme B expressed in the lymphocytes were detected by RT-PCR. The serum glucose, insulin and glucagons were also measured with routine methods. The pancreatic graft tissues were collected for pathologic examinations at the 1st, 3rd, 5th and 7th day after the operations.No differences in serum glucose, insulin and glucagons had been detected among the three groups (P0.05). The landraces in the transplantation plus immunosuppressant group had significantly lower expression of the mRNA of perforin and granzyme B in the lymphocytes and the local transplanted pancreas than those in the transplantation group (P0.05). The change of mRNA expression of perforin and granzyme B in the lymphocytes occurred 2-4 days earlier than the pathological changes when the acute rejection started.Monitoring the change of mRNA expression of perforin and granzyme B in the lymphocytes from the peripheral blood can help the early diagnosis of acute rejection of pancreatic transplantation.

To study the immune tolerance induced by human Ad-CTLA-41g after pancreaticoduodenal transplantation of rats.Wistar rats with diabetes were induced by caudal intravenous administration of streptozotocin (STZ) at a single dose of 60 mg/kg. Pancreaticoduodenal transplantations were performed with the SD rats as donors and the diabete Wistar rats as recipients. The SD pancreaticoduodenal grafts with human Ad-CTLA-41g infection via arterial perfusion in vitro were transplanted into Wistar rats with diabetes. The blood glucose of the rats was measured after transplantation. The graft functional survival time (GFST) was recorded. The concentration of IL-2 was measured by ELISA on day 3 and day 10 after operation. The human CTLA-41g was detected by Western blot after operation.The GFST of control group without gene transfer was (11+/- 1) d, while that of gene transfer group was (33 +/-4) d (P0. 01). The expression of serum human CTLA-41g was detected in rats of gene transfer group on the 10th day after transplantation, while no serum human CTLA-4Ig was detected in rats of control group without gene transfer. On the 3rd day after transplantation, the level of serum IL-2 was (33. 710+/-7. 803) pg/mL in rats of transfer group, and was (44. 772+/-9. 102) pg/mL in rats of control group without gene transfer. On the 10th day, the level of serum IL-2 was (47. 846+/-14. 050) pg/mL in rats of transfer group, and was (80. 806+/-18. 629) pg/mL in rats of control group without gene transfer. The levels of serum IL-2 in rats of transfer group was much lower than those of the control group without gene transfer (PO. 01).In vitro perfusion with human Ad-CTLA-41g is an effective method to transfer the target gene into pancreaticoduodenal graft. Ad-CTLA-41g infection can induce immune tolerance in rat with pancreaticoduodenal transplantation.

To explore the diagnosis and surgical treatment of abdominal cocoon.The clinical data of 16 patients with abdominal cocoon admitted to our hospital between Jun. 1993 and Oct. 2006 were analyzed retrospectively.Preoperatively, Barium meal X-rays revealed coils of intestine in 8 cases, incomplete intestinal obstruction in 6 cases, and prolonged intestinal transit time in 2 cases. CT scan showed dilated intestine and intestinal loops seemed to be encapsulated in a thickened capsule. After opening the peritoneum, entire or partial intestine encapsulated in thickened membrane encasing were found, including partial intestine encapsulated in 1 cases(Type I), entire intestine encapsulated in 2 cases (Type II), and entire intestine and other organs encapsulated in 12 cases(Type III). All the cases underwent adhesiolysis. Intestinal splint was done in 2 patients, gastrostomy in one patient with chronic pyloric obstruction, radical resection of rectal cancer in one patients and ileocolic resection in one patients with Crohn's disease. All patients were healed by surgical operation and confirmed the diagnosis histopathologically.Abdominal cocoon is rare. It is difficult to make a right diagnosis preoperatively. Barium meal X-rays and CT scan are useful methods for its diagnosis. For the treatment, attention should be paid on complete resection of fibrous membrane, adhesiolysis and prevent intestinal obstruction.

To evaluate the curative effect of curved cutter stapler (Contour, Ethicon Endo-Surgery, Inc) in the ultra low anterior resection for low rectal cancer.Clinic data of 56 patients with low rectal cancer from Dec. 2005 to Sep. 2006 were reviewed retrospectively. After total mesorectal excision (TME) and lateral lymph node dissection (LLD) in 56 cases, the rectal (anal) remnant was cut and closed with curved cutter stapler (Contour), and preserved for ultra low colo-rectal (anal) anastomoses with 33 mm straight intraluminal stapler.There was no operational death and the mean hospitalization time was (11.2+/-3.2) days. The incidence rate of postoperative complications in 1 month was 3.57% (2/65). Both of the cases were anastomotic leakage. One was cured by surgical drainage, the other combining with rectal vaginal fistula was cured by transverse colostomy.Curved cutter stapler has the advantages of complete cutting, safe closure and low anastomotic leakage rate in the process of ultra low anterior resection for low rectal cancer.

To clone and express the fused gene fragment coding rhoptry protein ROP2 and major surface protein P30 from Toxoplasma gondii as a preparation for the construction of the complex ROP2-P30 antigen by gene engineering.The gene fragment encoding P30 was amplified by PCR from T. gondii RH strain and subcloned into the recombinant plasmid pUC119/ROP2 already constructed. The recombinant plasmid pUC119/ROP2-P30 was digested by Sac I/HindIII and inserted into the same site of expression vector pET28b. The recombinant plasmid of pET28b/ROP2-P30 was transformed to E. coli and expressed under the induction of IPTG.The gene fragment 700 bp encoding P30 was obtained from the total DNA of T. gondii by PCR. The recombinant plasmid pET28b/ROP2-P30 was successfully constructed, which was highly expressed in E. coli, a fusion protein with molecular weight of 69000.The fusion gene encoding the rhoptry protein ROP2 and the major surface protein P30 of T. gondii has been successfully cloned and expressed to be an expected recombinant fusion protein ROP2-P30 with molecular weight 69000.

To develop a method for detecting the genotype of Plasmodium vivax merozoite surface protein 1 (PvMSP-1) alleles.According to the sequence characteristic of PvMSP-1, nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to amplify the polymorphic region of ICB5-ICB6 which contains Q repeats and PvuII restriction site (Sal-1 type). The PCR product was digested by PvuII restriction endonuclease and the digested fragments were observed by 2% agarose gel electrophoresis. The allelic type was determined according to the banding pattern.Bands in size of 400 bp (Belem type) and/or 470 bp (Sal-1 type) appeared in all 98 P. vivax isolates, no band was found in negative control. After PvuII digestion, two Sal-1 type fragments (120 bp and 350 bp) were obtained from 45 samples of 470 bp. Single-band of 400 bp appeared in 3 of 40 samples with 400 bp as Belem type, two bands of 120 bp and 280 bp appeared from other 35 samples as recombination type III, and another 2 bands with 120 bp and 240 bp as Korean isolate.The result showed that the nested PCR-RFLP may be applied in the detection and identification of the three PvMSP-1 allelic types in China.