ObjectiveTo investigate the expression of Twist and N-cadherin in pancreatic cancer,and to study its relationship with clinicopathological parameters and patients' prognosis.Methods The expression of the Twist and N-cadherin in 62 tissue samples from patients with pancreatic ductal adencocarcinoma and 10 normal pancreatic tissue samples was determined by using immunohistochemistry MaxVision two step method,and the relationship with clinicopathological parameters and patients' prognosis was analyzed.ResultsThe positive expression rate of Twist in pancreatic cancer was higher than that of normal pancreatic tissues (96.8％ vs.30％,P ＜0.01 ),and the positive expression rate of N-cadherin was higher than that of normal pancreatic tissues (75.8％ vs.0,P ＜0.01),but there was no correlation between them (r =0.100,P =0.441 ).The expressions of Twist and N-cadherin was significantly correlated with TNM stage,lymph node metastasis,infiltration of portal vein or nerves and tumor location ( P ＜0.05 ),but not with age,gender and degree of differentiation ( P ＞ 0.05 ).The post-operative survival of patients decreased with the increasing Twist expression,but the survival was not associated with the expression of N-cadherin.TNM stage,the expression of Twist was independent predictive factors of prognosis for pancreatic cancer patients.ConclusionsTwist and N-cadherin are highly expressed in pancreatic cancer,and the expression was associated malignant behavior of pancreatic cancer.The abnormal expression of Twist may be a potential marker for prognosis evaluation of pancreatic cancer patients.

Objective To investigate the effect of interferon-alpha (IFN-α) on preventing pancreatic fibrosis and expression of pancreatic α-smooth muscle actin (α-SMA) and collagen type m in experimental rat model induced by diethyldithiocarbamate (DDC). Methods Forty male Wistar rats were randomly divided into three groups: normal control group, fibrotic group(PF group) and IFN-α prevention group(IFN-α group). Pancreatic fibrosis were induced by repeated intraperitoneal DDC injection. IFN-α group were treated with subcutaneous injection of IFN-α in saline, administered daily at the dosage of 105 U. Six weeks later, all the rats were sacrificed, the samples of the pancreatic tissue obtained by biopsy were submitted to histological studies. The expression of α-SMA and collagen type Ⅲ in pancreatic tissues were detected by immunohistochemistry. Results From the 4th week on, the weight of rats in PF group grew slowly, some rats 'weight even decreased, while the weight of rats in IFN-α group kept growing up slowiy, and the difference became significant at the 5th week (306.17±20.89 g vs 279.17±23.58 g, P＜0.05 ). The fibrosis score, Masson stain score, α-SMA and pancreatic collagen type Ⅲ in PF group were 2.679±0.899, 218. 713± 36. 102, 148.971±30. 686 and 88. 142±42.581, respectively, and the corresponding values in PF group were 1.952±0.219, 114.732±24.912, 77.237±9.275 and 59.952±25.498, respectively. The extent of fibrosis in IFN-α group was significantly better than that in PF group (P＜0.05). Conclusions IFN-α may inhibit the extent of fibrosis, decrease the expression of α-SMA and pancreatic collagen type Ⅲ, therefore, it may partially prevent pancreatic fibrosis induced by DDC.

Objective To investigate the role of interleukin-18(IL-18) and interleukin-15(IL-15) in the mechanism of acute pancreatitis.Methods The serum levels of IL-18 and IL-15 were detected by enzyme-linked immunosorbent assay(ELISA) in SAP group(n=7) and MAP group(n=19).The relationship between IL-18 and IL-15 were analyzed.Results SAP group was compared with the MAP group:IL-18 (78.32?15.45)ng/L vs (28.21?13.18)ng/L,P

Objective To explore the value of pretargeting technology in vitro MRI of L5 peptide guided streptavidin-conjugated and polyethylene glycol modification protected ultra-small superparamagnetic iron oxide(SA-PEG-USPIO) to hepatocellular carcinoma(HCC) via glypican-3(GPC3) receptor.Methods Direct immumofluorescence assay with carboxyfluorescein(FAM) labeled L5 and competitive inhibition was performed in HepG2 and HL-7702 cells.Imaging was obtained from fluorescent microscope.Immunoassay fluorescence images were carried out to determine the expression of GPC3 in HepG2 cell.PEG-USPIO conjugated with streptavidin was made by carbodiimide reaction,and the hydrodynamic diameters,Zeta potential and magnetic relaxivity of SA-PEG-USPIO and PEG-USPIO were measured.HL7702 cells were used for evaluate cells viability of SA-PEG-USPIO and PEG-USPIO.HepG2 and HL-7702 cells were used as experimental and control group respectively.Each of the two cell lines were further divided into three groups:L5-BT united SA-PEG-USPIO group,SA-PEG-USPIO group and control group.Prussian blue staining and MRI was preformed to observe the targeting efficacy of SA-PEG-USPIO respectively,and normalized T2 signal was recorded.The significant changes of normalized T2 signal intensity among groups was deterumine by using One-way analysis of variance.Results There were much more fluorescences on the membrane and cytoplasm of HepG2 cells than those on HL-7702 cells and cells of competition group.And indirect immunofluorescence images show the obvious expression of GPC3 in HepG2 cell.The SA-PEG-USPIO and PEG-USPIO nanoparticles had hydrodynamic diameters of (22.73 ± 3.31) and (35.97±5.19)nm,Zeta potential of them were (4.22±0.53) and (-7.91± 1.22)mV and magnetic relaxivity were 0.139 4× 103 and 0.103 9 × 103 mM-1s1.Although the highest concentration of SA-PEG-USPIO and PEG-USPIO was 2.4 mmol/L,cells viability was greater than 80％.The most iron particle was observed in L5-BT united SA-PEG-USPIO group of HepG2 cells.In vitro MR,the normalized T2 signal intensity of HepG2 cells in L5-BT united SA-PEG-USPIO group,SA-PEG-USPIO group and control group were 39±7,77 ± 12 and 93 ± 4.There was significant difference among those three groups (F=23.96,P＜0.01).The normalized T2 signal intensity of HL-7702 cells in each of three groups were 69± 11,78±8 and 95±5.There was no significant difference among those three groups (F=2.86,P＞0.05).Conclusion By the pretargeting method,L5 peptide guided SA-PEG-USPIO has effective targeting ability to HepG2 cells in vitro.

Objective To investigate the mechanism of mastoparan-1 (MP-1) antagonizing lipopolysaecharide (LPS) in vitro. Methods The affinity of MP-1 for lipid A was assayed by biosensor, and the neutralization of MP-1 on LPS (2 μg/L) was detected by kinetic turbidimetric limulus test. After exposing fluorescin isothiecyanate (FITC) labeled LPS (FITC-LPS) to MP-1 at different concentrations (5, 10, 20, 40 μmol/L), the binding of FITC-LPS to murine RAW264.7 cells was analyzed by laser scanning confocal microscopy. The influence of MP-1 on TLR4 expression in RAW264.7 cells stimulated by LPS (100 μg/L) was detected by immunoeytochemieal staining. The expressions of TLR4, TNF-α and IL-6 at the gene and protein level were detected by RT-PCR and ELISA after exposing LPS (100 μg/ L) stimulated RAW264.7 cells to MP-1 at different concentrations. The effect of MP-1 on the viability of RAW264.7 cells was detected by MTT assay. Results MP-1 had high affinity to lipid A and could neutralize LPS. MP-1 at 10 μmol/L significantly inhibited not only binding of FITC-LPS to RAW264.7 (P < 0.05), but also protein and gene expressions of TLR4, TNF-α and IL-6 in LPS stimulated RAW264.7 cells in a dose-dependent manner (P < 0.05). No toxic effect of MP-1 on the viability of RAW264.7 cells was found (P > 0.05). Conclusions MP-1 inhibits cell viability mediated by LPS, which may be related to its neutralization of LPS and inhibition of binding of LPS to RAW264.7 cell membrane receptors.

Objective To study the serotype , biochemical characteristics , virulence gene and multilocus sequence typ-ing(MLST) of S.flexneri 4c in Beijing, Shanghai and Shenyang .Methods Seventy-six strains of S.flexneri 4c isolated from stool samples which had been collected from above-mentioned cities of China were identified with Denka Seiken serum and MASF monoclonal serum .Biochemical characteristics of each strain were identified by API 20E test strip and PCR technology was used for detecting 12 pair virulence genes of S.flexneri.MLST was used to analyze the characteristics . Results The serum agglutination antigen structure of S.flexneri 4c was(Ⅳ:7,8).MASF:B+,Ⅳ:Ⅰ+,7 (8) +.S.flexneri 4c developed different results in biochemical reactions and carried different rates of virulence genes , respectively .The IND test positive rate was 17.11%; MEL weakly positive rate was 3.9%, and ARA test weakly positive rate was 22.37%. Virulence genes were carried at a rate of 89.47% -100%, MLST typing was ST245.Conclusion S.flexneri 4c with serum agglutination antigen structure (Ⅳ:7,8) is a new serotype of S.flexneri.The main biochemical reactions are glucose fermentation and mannitol decomposition .A variety of Shigella related virulence genes are carried .MLST generation is consistent,suggesting that the bacteria might have evolved from ST 270 cloning.

OBJECTIVETo investigate the application of tourniquet in burn patients during tangential excision on the extremities.

METHODSSeventy - nine burn patients who were arranged to receive tangential excision and skin grafting on the extremities were randomly divided into A and B groups. The patients in A group (n = 41) underwent the operation with the tourniquet applied continuously throughout the operation, while those in B group (n = 38), only with tourniquet applied during tangential excision. The amounts of blood loss and blood transfusion, the operation time and the take rate of grafted skin and the incidence of complications were investigated and recorded.

RESULTSThe amounts of blood loss and blood transfusion during operation in A group were 42% and 50% less than those in B group, respectively (P < 0.001). Moreover, the operation time on the upper and lower extremities in A group was much shorter (for 41% and 37%, respectively) than those in B group (P < 0.001). In addition, there was no difference of the take rate of skin graft and the incidence of subcutaneous hematoma between the two groups (P > 0.05).

CONCLUSIONContinuous tourniquet application during tangential excision on the extremities in burn patients was proved to be effective in reducing operational blood loss, blood transfusion and in shortening operation time.

Adult ; Blood Loss, Surgical ; prevention & control ; Blood Transfusion ; Burns ; surgery ; Extremities ; surgery ; Humans ; Skin Transplantation ; Tourniquets

Objective To explore the effect of JianpiYiqi Decoction on the proliferation,invasion and apoptosis of Human hepatoma Huh-7 cells under the constructed inflammatory microenvironment stimulated by lipopolysaccharide(LPS)and adenosine triphosphate(ATP),and to detect the interleukin-1β(IL-1β)and interleukin-18(IL-18)Expression level in cells.Methods Construction drug-containing serum of JianpiYiqi.Adopt LPS and ATP were added to the cell culture medium to stimulate hepatoma cells and construct the inflammatory microenvironment.The cells were divided into blank group,model group,VX-765 group(10 μmol·L-1),low concentration of traditional Chinese medicine group(15%JianpiYiqi Decoction low dosedrug-containing serum),Medium concentration of traditional Chinese medicine group(15%JianpiYiqi Decoction Medium dose drug-containing serum),and high concentration of traditional Chinese medicine group(15%JianpiYiqi Decoction high dose drug-containing serum).Adopt CCK-8 method was used to detect the proliferation level of cells in each group after cell intervention;The apoptosis rate of cells in each group was detected by Annexin V-FITC/PI method;Adopt Transwell method was used to detect the level of cell invasion in each group;adopt PI single staining was used to detect the cell cycle level in each group;Adopt ELISA method and Western blot method were used to detection of IL-1β and 1l-18 expression level in Huh-7 cells.Results Compared with the blank group,in the model group stimulated by LPS and ATP,the proliferation level,invasion level,IL-1β and IL-18 expression level of Huh-7 cells were higher(P<0.05),and the apoptosis level was lower(P<0.05).Compared with other groups,the Medium concentration of traditional Chinese medicine group and the high concentration of traditional Chinese medicine group could effectively inhibit the proliferation and invasion level of Huh-7 cells,block the cell proliferation cycle,reduce the cell survival rate(P<0.05),significantly induce apoptosis of Huh-7 cells(P<0.05),and reduce the expression levels of IL-1β and IL-18 in Huh-7 cells(P<0.05).Conclusion The drug-containing serum JianpiYiqi.The medium and high concentrations of JianpiYiqi Decoction drug-containing serum can inhibit the proliferation and invasion,block the cell cycle and induce apoptosis of human liver cancer Huh-7 cells by improving the inflammatory microenvironment in liver cancer Huh-7 cells.Its mechanism may be related to the inhibition of liver cancer cells.Analyze the levels of inflammatory factors IL-1β and IL-18 to achieve the therapeutic purpose of primary liver cancer.

Objective To investigate the effect ofcranioplasty on prognosis of patients accepted intracranial aneurysm clipping combined with simultaneous decompressive craniectomy.Methods One hundred and forty-four patients accepted intracranial aneurysm clipping combined with simultaneous decompressive craniectomy (first-stage operation) in our hospital from January 2013 to June 2017 were chosen;there were 56 patients without cranioplasty in the control group and 88 patients with cranioplasty (second-stage operation) in the observation group.The degrees of coma before first-stage operation were assessed by Glasgow coma scale (GCS).The general state three months after first-stage operation was assessed by GCS and activity of daily living (ADL) scale.The prognoses of these patients 9 and 15 months after first-stage operation were assessed by Glasgow outcome scale (GOS) and ADL scale.The clinical data,prognoses and incidence of hydrocephalus of patients from the two groups were compared.Related factors associated with hydrocephalus were analyzed by multivariate Logistic regression analysis.Results GCS,GOS and ADL scale scores in the observation group 9 and 15 months after first-stage operation were all significantly higher than those in the control group (P＜0.05);incidence of hydrocephalus in the observation group after first-stage operation (31.82％) was significantly lower than that in the control group (62.5％,P＜0.05).Logistic regression model revealed that cranioplasty,Hunt-Hess grading and Fisher grading were independent related factors for incidence of hydrocephalus (P＜0.05);cranioplasty was the protective factor of hydrocephalus (OR=0.126),and Hunt-Hess grading and Fisher grading were the risk factors of hydrocephalus (OR=5.311 and 5.073).Conclusion Cranioplasty can reduce the incidence of hydrocephalus and improve the prognosis of patients accepted intracranial aneurysm clipping combined with simultaneous decompressive craniectomy.