Objective To study the changes of endothelin-1(ET-1) and calcitonin gene-related peptide(CGRP) in plasma of cerebral vasospasm(CVS) after resection of skull base tumors and the effect of the two factors on cerebral vasospasm. Methods Totally 34 cases were divided into symptomatic cerebral vasospasm group,asymptomatic cerebral vasospasm group and nonvasospasm group after resection of skull base tumors.The blood specimens were obtained from the 34 patients on days 1,3,5,7 and 14 after the resection.The concentration of ET-1 and CGRP was detected by radioimmunoassay;meanwhile,transcranial doppler was recorded.Another 10 normal adult served as control group. Results ① Concentration of ET-1 in plasma elevated from the 1st day after resection of skull base tumors,reaching peak levels on day 5 to day 7,then decreased gradually and nearly recoverd on day 14.Concentration of CGRP in plasma decreased from day 3 after resection of skull base tumors,with the lowest concentration on day 7,then increased gradually and recoverd on day 14.② Concentration of ET-1 in plasma of the three groups was higher than that of normal adult group,while concentration of CGRP of the three groups was lower than that of normal adult group.③ Concentration of ET-1 in plasma in vasospasm groups was higher than that in nonvasospasm group(P

Objective To construct a Polyphosphate kinase 1 ( ppk1) gene deletion mutant of uro-pathogenic Escherichia coli (E.coli) CFT073, and to explore the biological properties of the mutant strain . Methods The ppk1 gene deletion strain (△pk1) was constructed based on CFT073 E.coli strain by usingλRed homologous recombination technology .A comparison analysis was conducted on adhesive and invasive abilities between CFT073 wild type strain and △pk1 strain in in vitro model of human bladder cancer epithe-lial cell 5637 .Crystal violet staining method was used to evaluate the influences of ppk1 gene deletion on biofilm formation.Results The CFT073 ppk1 deletion mutant strain was successfully constructed .Com-pared with the wild type strain ,△pk1 strain showed impaired adhesive and invasive abilities to 5637 cells. Moreover , the absorbance values of crystal violet at 570 nm at each time point of the mutant strain group were also lower than those of the wild-type strain group .Conclusion The ppk1 gene deletion mutant of uro-pathogenic E.coli CFT073 could be successfully constructed by Red homologous recombination technology and its biological properties indicates that ppk1 gene plays an important role in the pathogenesis of uropatho-genic E.coli infection through regulating the abilities of adhesion , invasion and biofilm formation .

Objective To explore the role of ppk1 gene in E.coli CFT073 strain during urinary tract infection (UTI). Methods C57BL/6 mouse models of acute UTI with the wild-type(CFT073) and ppk1 mutant (△pk1) infected, were used to compare the bacteria colonization and inflammation induction abilities of bladder tissues. Results In the mouse models, the △pk1 strain showed a significantly lower infection rate (73.3% vs 93.3%) and lower adhesion frequency of bladder (0.01%vs 0.5%) than those of the CFT073 strain. The expression of IL-6 and TNF-αwere reduced in the bladder of △pk1 infected group (P<0.05). Hematoxylin-Eosin tissue staining showed that the damage degree of bladders in △pk1 infected mice were less serious than the CFT073 infected mice. Conclusion ppk1 gene plays an important role in E.coli colonization to bladder and the inflammation induction ability.

Objective: To examine the expression of glucocorticoid receptor in human colon cancinoma tis-sues and call lines and to explore the survival of colon cancer cell lines treated with dexamethasone alone or in combination with 5-Fu and L-OHP in a way of dexamethasone pretreatment or co-administration in vitro.Methods: The expression of glucocorticoid receptor was detected in 61 cases of colon cancer tissue samples and 4 types of colon cancer cell lines by immunohistochemistry. Apoptosis was detected by Hoechst33342 staining and flow cytometry. MTT assay was employed to detect the chemosensitivity of colon carcinoma cells to L-OHP and 5-Fu with dexamethasone pretreatment for 24 hours or co-administretion. Results: Positive GR expression was found in 57.3% colon cancer tissue samples and in Lovo and HCT-116 cell lines, not in HT-29 and SW-480. Apoptosis was detected in GR-expressed Lovo and HCT-116 cells at 72 hours after 1×10~(-4)mol/L Dex treatment, and the rates of apoptosis were higher than those in the control groups without Dex (P<0.01),GR-negative cells, HT-29 and SW-480 even treated with 1 × 10~(-4)mol/L Dex for 72 hours. Pretreatment and co-administration for Lovo cells with 1×10~(-4)mol/L Dex could decrease the IC50 of L-OHP from 13.7±1:1.3μg/mL to 5.9±0.6μg/mL and 4.8±0.7μg/mL, respectively. IC50 of 5-Fu was decreased from 72.2±8.1 μg/mL to 21.1±4.1μg/mL and 18.6±4.0μg/mL, respectively. Conclusion: There is expression of glucocorticoid receptor in part of colon carcinoma tissue samples and cell lines. Apoptosis does occur in GR-expressed Lovo and HCT-116 cells induced by dexamethasone in vitro. Pretreatment for 24h and co-administration with Dex can increase the chemosensitivity of Lovo cells to L-OHP and 5-Fu.

Objective To investigate the role and the mechanism of ppk1 gene (coding for polyphosphate kinase 1) in oxidative stress resistance in uropathogenic Escherichia coli (UPEC).MethodsMutant strains with ppk1-deletion (△pk1) and complemented strains (△pk1-C) were constructed based on the UPEC strain CFT073.A comparative analysis was conducted to analyze survival rates of CFT073, △pk1 and △pk1-C strains at different time points while they were under oxidative stress.Differences in protein expression between CFT073 and △pk1 strains were analyzed using mass spectrometric analysis.Differences between CFT073 and △pk1 strains in expression of katG and katE genes were analyzed using real-time quantitative RT-PCR.Results The survival rate of △pk1 strains was lower than that of CFT073 strains at every time point, while the survival rate of △pk1-C strains was basically the same as that of CFT073 strains.Gel image analysis and mass spectrometric analysis revealed that six proteins were down-regulated and one was up-regulated in △pk1 strains as compared with those in CFT073 strains.Expression of the catalase-coding genes katG and katE in △pk1 strains were respectively (20.5±8.2)% and (20.9±6.9)% of those in CFT073 strains (P<0.05).Conclusion The ppk1 gene plays an important role in oxidative stress resistance in UPEC by modulating the expression of catalase-coding genes katG and katE.

OBJECTIVETo investigate the optimal reconstruction technique after total gastrectomy.

METHODSA total of 159 patients with gastric cancer undergoing total gastrectomy in Tianjin Cancer Hospital between January 2005 and December 2007 were divided into 4 groups according to the reconstruction technique: group A(functional jejunal interposition with a pouch, n=46), group B(modified Braun type II(, n=38), group C (P pouch with Roux-en-Y esophagojejunostomy, n=25), group D(Roux-en-Y esophagojejunostomy, n=50). Quality of life(QOL), nutritional status 1 year after surgery, and perioperative complications were analyzed.

RESULTSThere were no significant differences in perioperative complications(P>0.05). One year after operation, QOL(Visick index) was better in group A than that in group B, C and D(P<0.05), and group D was inferior to group A, B and C(P<0.05). The increase in food intake, weight gain, hemoglobin and total protein were better in group A than those in group B, C and D(P<0.05) and group D was inferior to group A, B and C(P<0.05). The prognostic nutrition index ratio of the four groups were 1.21±0.15, 1.14±0.97, 1.15±0.16, and 1.10±0.16, respectively. Group A was better than that in group B, C and D (P<0.05) and group D was inferior to group A, B and C(P<0.05). The incidences of dumping syndrome, reflux esophagitis, Roux-en-Y stasis syndrome in group A were 4.3%(2/46), 2.2%(1/46) and 2.2%(1/46), respectively, which were significantly lower than those in other groups (P<0.05).

CONCLUSIONSFunctional jejunal interposition with a pouch is associated with improved nutritional condition and quality of life, and less perioperative complications. It is a reasonable reconstruction method after total gastrectomy.

Adult ; Aged ; Anastomosis, Roux-en-Y ; methods ; Anastomosis, Surgical ; methods ; Esophagus ; surgery ; Female ; Follow-Up Studies ; Gastrectomy ; methods ; Humans ; Jejunum ; surgery ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Retrospective Studies ; Stomach Neoplasms ; surgery ; Treatment Outcome

Objective To study the FMS-like tyrosine kinase-3 (FLT3) gene, NPM1 gene and c-kit gene mutations in acute myeloid leukemia (AML) by extracting DNA from the storage of bone marrow slides, and to investigate the relationship between the three gene mutations and clinical features in AML. Methods The bone marrow slides of 55 patients diagnosed with AML were enrolled in this study. The PCR, DNA sequencing and molecular cloning were used to detect and analyse the FLT3-ITD, NPM1 and c-kit gene mutations. Patients' remission, progression and survival time were also recorded. Results The DNA was successfully extracted from the bone marrow slides with -20 ℃ frozen storage without Wright stained, chemically fixed, and room temperature storage Wright stained discoloured by phenol ∶ chloroform ∶ isoamyl alcohol method, which can be used in PCR, direct sequencing and molecular cloning sequencing analysis. 10 of the 55 cases (18.2 %) were FLT3-ITD positive, including 9 cases with heterozygous mutations and 1 case with homozygous mutation. FLT3-ITD positive group had lower complete remission (CR) rate, shorter event-free survival (EFS) time and overall survival (OS) time than the negative group (P< 0.05). 9 of the 55 cases (16.4 %) had NPM1 heterozygous gene mutations, all belonging to type A. The EFS rate of the patients with NPM1 mutation was higher in 10 months and the OS rate was higher in 19 months (P< 0.05). 3 of 9 NPM1 mutations patients were FLT3-ITD positive. The CR rates of the four groups after initial remission induction therapy in order were NPM1+FLT3-ITD-, NPM1-FLT3-ITD-, NPM1-FLT3-ITD+, NPM1+FLT3-ITD+(P<0.05). Besides, NPM1-FLT3-ITD+was a risk factor affecting the OS (RR=1.250, P=0.005). 2 of the 55 cases (3.6 %) had c-kit gene mutations, namely mutant D816H and mutant D816V. The c-kit gene mutations were not found in patients with FLT3-ITD and NPM1 mutations. Conclusions The FLT3-ITD mutation is a poor prognosis molecular marker in AML, and NPM1 mutation is a good factor for the prognosis. NPM1-FLT3-ITD+is a risk factor affecting OS. The rate of c-kit gene mutation is low in AML, without the overlap of FLT3 and NPM1 mutations.

Objective To investigate the expression levels of serum LINC00978 in gastric cancer patients and its correlation with clinicopathological characteristics,and evaluate its clinical diagnostic value.Methods The expression levels of LINC00978 in gastric cancer cells and serum samples from the patients with gastric cancer or gastric benign diseases and healthy controls were detected by real-time qRT-PCR,and its correlations with clinicopathological parameters were analyzed.The diagnostic efficacy of serum LINC00978 was evaluated by the receiver operating characteristic (ROC) curve.Results The expression levels of LINC00978 in MGC-803 cells were significantly higher than that in GES-1 cells (18.88 ± 1.15 vs 1.00 ± 0.03,t =-21.926,P ＜ 0.05).The expression levels (median[P25,P75]) of serum LINC00978 in gastric cancer patients (3.525 [1.385,8.954]) were significantly higher than those in the patients with gastric benign diseases (0.419 [0.258,1.369],Z =-4.834,P ＜ 0.01) and healthy controls (0.814 [0.351,2.510],Z =-4.686,P ＜ 0.01).The expression levels of serum LINC00978 in gastric cancer patients were significantly related to lymphatic metastasis (r =0.448,P ＜ 0.01),invasive depth (r =0.369,P ＜ 0.01) and TNM stage (r =0.383,P ＜ 0.01),and those after operation significantly lower than before operation (0.17 [0.15,0.39] vs 0.98 [0.59,1.61],Z =-5.731,P ＜ 0.01).There was no significant difference in serum LINC00978 levels between the patients with gastric benign diseases and healthy controls (Z =-1.693,P ＞ 0.05).The area under the ROC curve (AUCROC) and 95％ confidence interval (CI) of serum LINC00978 levels in the diagnosis of gastric cancer were 0.807 and 0.732-0.882,respectively.When the cutoff value was 1.806,the sensitivity and specificity for the diagnosis of gastric cancer were 71.4％ and 75.9％,respectively.Conclusion The expression levels of serum LINC00978 in gastric cancer patients increase and are related to lymphatic metastasis,invasive depth and TNM stage,which may be served as a novel biomarker for the diagnosis of gastric cancer.

Objective To detect the expression levels of anti-differentiation antagonizing noncoding RNA (DANCR) in exosomes from serum of gastric cancer patients and evaluate its clinical application.Methods The expression levels of DANCR in exosomes derived from the culture supernatants of gastric cancer cell lines and the serum samples of gastric cancer patients were detected by using realtime qRT-PCR.The correlations between DANCR levels and the clinicopathological parameters were analyzed.A receiver operating characteristic(ROC) curve was drawn to evaluate the diagnostic efficiency of DANCR.Results The expression level of DANCR in HGC-27 cell(4.43 ± 0.37) was significantly higher than that in GES-1 cells (1.01 ± 0.01) (P ＜ 0.05).The expression levels of DANCR in exosomes from the supernatants of HGC-27 cells(3.06 ±0.14) were significantly higher than that from GES-1 cells(1.01 ± 0.20) (P ＜ 0.05).The expression levels of DANCR in exosomes from the serum samples of gastric cancer patients [5.060 (1.380,22.785)] were significantly higher than that in gastric benign disease [0.535 (0.340,1.380)] (Z =-3.409,P ＜ 0.01) and healthy controls [1.200 (0.605,1.655)] (Z =-4.229,P ＜ 0.01).There was no significant difference in the expression levels of serum exosomal DANCR between gastric benign disease group and healthy control group(Z=-1.308,P ＞ 0.05).In addition,the expression levels of DANCR in gastric cancer patients were related to tumor size,TNA stage and lymph node metastasis.The area under the ROC curve of serum exosomal DANCR expression levels was 0.777 in the diagnosis of gastric cancer,the 95％ confidence interval was 0.678 to 0.876 and the cutoff value was 2.50.The sensitivity was 68.6％ and the specificity was 84.4％.Conclusion The expression level of serum exosomal DANCR may increase in gastric cancer patients and could be served as a novel marker for the diagnosis of gastric cancer.

OBJECTIVETo analyze the clinical and pathologic influencing factors of early recurrence in patents with gastric cancer after radical gastrectomy.

METHODSClinicopathological data of 141 patients with recurrence after curative gastrectomy for gastric cancer from January 2001 to December 2004 were analyzed retrospectively. Risk factors correlated with tumor early recurrence and survival difference between early recurrence group (< 1 year, 82 cases) and control group (1 year after, 59 cases) were assessed.

RESULTSThe 1- and 3-year survival rates of in early recurrence group and control group were 36.6%, 2.4% and 100%, 45.8%, respectively (P < 0.05). The median survival time after recurrence in the two groups was 3, 5 months, respectively (P < 0.05). Univariate analysis showed that the age, tumor Borrmann type, tumor site, invasive depth, lymph node metastasis, pTNM stage, metastatic lymph node ratio, surgical procedure and intraperitoneal hyperthermic perfusion chemotherapy (IHPC) were significant factors associated with early recurrence after curative gastrectomy for gastric cancer (P < 0.05). Lymph node metastasis, metastatic lymph node ratio and IHPC were independent factors associate with early recurrence after curative gastrectomy on multivariate analysis (P < 0.05).

CONCLUSIONSThe patients with early recurrence after the radical gastrectomy have a poorer survival compared with cases recur later. Lymph node metastasis, metastatic lymph node ratio and IHPC are independent factors associate with early recurrence after curative gastrectomy for gastric cancer.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Gastrectomy ; Humans ; Kaplan-Meier Estimate ; Logistic Models ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; etiology ; pathology ; Postoperative Period ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms ; pathology ; surgery