Objective To evaluate the role of pretreatment with docosahexaenoic acid (DHA) in the protective effect of angiopoietin-1 (Ang-1) on rat brain microvascular endothelial cells (BMVECs) during oxygen glucose deprivation (OGD).Methods BMVECs were sub-cultured in vitro and divided with a random number table into 7 groups:normal control group,normal control + Ang-1 group,OGD group,OGD + Ang-1 group,OGD + DHA group,OGD + DHA + Ang-1 group,and OGD + DHA + GW9662 + Ang-1 group.The normal control and normal control + Ang-1 groups were cultured in DMEM containing serum,5 mmol/L glucose,and 1.25 mmol/L pyruvate;OGD groups were cultured in glucose-and serum-free DMEM.DHA (40 μmol/L) was added to OGD + DHA,OGD + DHA + Ang-1,and OGD + DHA + GW9662 + Ang-1 groups,and 5 μmol/LGW9662 [inhibitor of peroxisome proliferator-activated receptor gamma (PPAR-γ)] to OGD + DHA + GW9662 +Ang-1 group,before pretreatment for 1 hour in 5％ CO2 and 95％ air.After the pretreatment,Ang-1 (250 ng/ml)was added to normal control + Ang-1,OGD + Ang-1,OGD + DHA + Ang-1,and OGD + DHA + GW9662 +Ang-1 groups.The cells were cultured in 94％ N2∶ 5％ CO2∶ 1％ O2 for 24 hours,except for normal control and normal control + Ang-1 groups,which were cultured in 5％ CO2 and 95％ air instead.The cell apoptosis rate was detected with flow cytometry,expressions of Bax,Bcl-2,caspase-3,Tie-1,Tie-2,pTie-2,pAkt,ZO-1proteins with Western blot.Results Compared with normal control group,the cell apoptosis rate in OGD,OGD + Ang-1,OGD + DHA,OGD + DHA + Ang-1,and OGD + DHA + GW9662 + Ang-1 groups were significantly increased (P =0.000,0.000,0.000,0.004,0.000);the expression levels of Bax,caspase-3,and Tie-1 were significantly enhanced (Bax:0.62 ± 0.03,0.38 ± 0.03,0.45 ± 0.03,0.26 ± 0.02,0.33 ± 0.02vs.0.16 ±0.01;caspase-3:0.76 ±0.05,0.42 ±0.04,0.52 ±0.02,0.32 ±0.02,0.40 ±0.02 vs.0.15 ±0.01;Tie-1:0.51 ±0.03,0.25 ±0.01,0.33 ±0.02,0.16±0.01,0.22±0.02 vs.0.12 ±0.01;all P=0.000);the expression levels of Bcl-2,Bcl-2/Bax,Tie-2,and pTie-2 were significantly decreased [Bcl-2:0.09±0.01,0.20±0.01,0.16±0.02,0.31±0.01,0.22±0.01 vs.0.34±0.01;Bcl-2/Bax:(14.93±1.86)％,(68.03±5.56)％,(36.93 ±2.22)％,(119.1 ±13.3)％,(64.23 ±6.07)％ vs.(208.33 ±7.37)％;Tie-2:0.07±0.01,0.16±0.02,0.11 ±0.01,0.21±0.01,0.18±0.01 vs.0.26±0.01;pTie-2:0.05±0.01,0.15 ±0.01,0.07 ±0.01,0.22 ±0.02,0.16±0.01 vs.0.27 ±0.01;allP=0.000],in addition,pAkt and ZO-1 in OGD,OGD + Ang-1,OGD + DHA,and OGD + DHA + GW9662 +Ang-1 groups were also significantly reduced (pAkt:0.13 ±0.01,0.26 ±0.01,0.14 ±0.01,0.28 ±0.02vs.0.39±0.02;ZO-1:0.08±0.01,0.18±0.01,0.10±0.01,0.19±0.01vs.0.23±0.02;allP=0.000).Compared with the OGD group,OGD + Ang-1 and OGD + DHA + Ang-1 groups demonstrated significantly reduced cell apoptosis (both P =0.000).Compared with normal control + Ang-1 group,the expression levels of pTie-2,pAkt,and ZO-1 were significantly decreased in OGD + Ang-1 and OGD + DHA +Ang-1 groups (pTie-2:0.15 ±0.01,0.22 ±0.02 vs.0.52 ±0.02;pAkt:0.26 ±0.01,0.37 ±0.04 vs.0.67 ± 0.05;ZO-1:0.18 ± 0.01,0.24 ± 0.02 vs.0.39 ± 0.05;all P =0.000).Compared with OGD +Ang-1 group,OGD + DHA + Ang-1 group had significantly decreased cell apoptosis and expression levels of Bax,caspase-3 and Tie-1,and significantly increased expressions of Bcl-2,Bcl-2/Bax,Tie-2,pTie-2,pAkt,and ZO-1 (all P =0.000),while OGD + DHA + GW9662 + Ang-1 group showed no significant differences in these indexes (P =0.202,0.770,0.382,0.448,0.233,0.736,0.143,0.526,0.495,0.670).Conclusion Pretreatment with DHA may enhance the protective effect of Ang-1 on rat BMVECs under the condition of OGD,possibly via activating PPAR-γ,suppressing Tie-1 expression,and hence amplifying the activation of Tie-2.

Objective To study the correlation between risk factors of CVD and cognitive deficit associated with cerebral infarction. Methods Cognitive function was tested in 101 cases of cerebral infarction using HDS-R at the end of 3 weeks and 6 months after onset of the stroke. Meanwhile, the data of risk factors and relative information of cerebral vascular disease were collected by using the scale of complications and inquiring the past history. The risk factors of CVD, such as neurological deficit, diabetes, hypertension, obesity, fever, heart diseases, smoking, overdrinking were compared between the cognitive deficit group and non-cognitive deficit group. Logistic regression analysis was adopted for data analysis. Results Of the 101 cases , 65 had cognitive deficit at the end of 3 weeks and 53 at the end of 6 months after onset of stroke. Age and score of scale of complications between cognitive deficit group and non cognitive deficit group were significantly different(P

Objective To evaluate the clinical efficacy and safety of endoscopic mucosal resection (EMR) assisted with magnifying chromoendoscopy in treatment of colorectal neoplasms. Methods Patients who met criteria for EMR including appropriate flat or depressed type and sessile lesions were enrolled. The association of morphology of colorectal lesions with histopathology was observed and the accuracy of estimation of invasive depth by magnifying chromoendoscopy was evaluated. Results Ninety lesions of 81 patients were reseeted by EMR (25 being sessile and 65 being flat or depressed). The histological results revealed low-grade dysplasia (LGD) in 58 lesions, high-grade dysplasia (HGD) in 20 lesions, and adenocarcinoma in 12 lesions. The average size of lesions was (1.4±0.5) cm in HGD, (1.6±0.5) cm in cancer and (1.0±0.4) cm in LGD with no significant difference (P> 0.05). It was shown that the flat and depressed lesions were more likely to be HGD or cancer as compared to sessile lesions, but with no statistical difference [41.5 % (27/65)vs. 20.0% (5/25), P= 0.084]. Moreover, the lesion with central depression was more likely to be HGD or cancer as compared to those without depressed surface [51.0% (25/49) vs. 17.1 % (7/41), P<0.01)]. The accuracy of estimating invasive depth by magnifying chromoendoscopy was 97.8% (86/90). Complete resection was confirmed histologically in 95.8% (88/90) of all lesions. Conclusions Colorectal lesions of depressed and flat types with central depression are more likely to be malignant. Estimation of invasive depth of colorectal neoplasia by magnifying chromoendoscopy in EMR treatment makes it more effective and safer.

Objective To evaluate the accuracy of preoperative three-dimensional endoscopic ultra-sonography (3D-EUS) in tumor invasion depth (T-staging) of rectal cancer. Methods From May 2007 to November 2007, 21 patients with rectal cancer diagnosed by eolonoscopy and biopsy, underwent 3D-EUS, 2D-EUS and MRI before surgery to predict T-staging. The results were assessed according to post-surgical pathological findings. Results The accuracy of 3D-EUS in T-staging was 85.71%, which was significantly higher than that of MRI (57.14%, P<0.05), and similar with that of 2D-EUS (76.19%, P>0.05). Conclusion More accurate T-staging could be achieved by 3D-EUS, which could be combined with MRI to detect lymph node metastasis, and provide more information for therapy decision in rectal cancer.

Objective To explore the feasibility,efficacy and safety of endoscopic submucosal dissection (ESD) for tumors originating from gastric muscularis propria.Methods A total of 20 patients with tumors originating from gastric muscularis propria confirmed by EUS and CT scan underwent ESD.Laparoscopic intervention was applied in 3 cases when difficulties in en bloc removal or perforation occurred.Results The mean ESD procedure time was 74.8 min (40-120 min),and the mean resected tumor size was 2.6 cm (1.5-3.5 cm).No severe complication occurred.Pathological findings were 17 cases of gastrointestinal stromal tumors and 3 cases of leiomyoma.Conclusion ESD is an effective method for treating the tumors originating from gastric muscularis propria,and laparoscopic intervention is necessary for en bloc resection in some cases.

Objective To evaluate the therapeutic effect of endoscopy for patients with anastomotic or nonanastomotic biliary strictures after liver transplantation.Methods Clinical and follow-up data of 56 patients,38 (67.9％) with anastomotic strictures (AS),and 18 (32.1％) with nonanastomotic strictures (NAS) after liver transplantation,who underwent endoscopic therapy were collected and analyzed.Results Compared to AS patients,NAS patients showed shorter time from liver transplantation to initial presentation (4.45 ± 1.47 vs.8.00 ±2.31 months,P =0.000),underwent more ERCP procedures (6.20 ±1.28 vs.4.11 ± 1.51,P =0.000) and received more stents (10.20 ±3.59 vs.7.53 ±2.12,P =0.001).Although the success rate was not statistically different,patients with NAS needed longer treatment duration and had higher recurrence rate (50.0％ vs.18.2％,P =0.033) as well as shorter duration from treatment to recurrence.Among patients with AS,the treatment failure group had longer stricture length (13.00 ±3.61 vs.6.63 ±2.09,P =0.000) than the success group and the patients who experienced recurrence had longer stricture length (10.48 ± 1.07 vs.5.86 ± 1.55,P =0.000) and narrower stricture diameter (1.52 ± 0.38 vs.1.94 ± 0.32,P =0.001).Among NAS patients,the multiple biliary strictures group had lower success rate and higher recurrence rate,but with no statistical significance (62.5％ vs.90.0％,P =0.275 and 66.7％ vs.37.5％,P =0.592,recpectively).Conclusion NAS patients have lower response to endoscopic treatment than AS patients.The length and diameter of biliary stricture in AS as well as the number of stricture in NAS may influent the effect of endoscopic treatment.

BACKGROUND: Permanent or transient implantation of biomaterials can result in biomaterials-centered infections (BCI) in lung cancer patients.OBJECTIVE: To investigate the relationship between BCI and peripheral blood transforming growth factor β1 (TGF-β1) in patients with lung cancer.METHODS: A total of 248 lung cancer patients undergoing in vivo intravascular catheter indwelling > 7 days were included.Quantitative method was used for intubation, bacteriological culture and paired blood culture, and API Staph strips were adopted for positive patients. While enzyme-linked immunosorbent assay was used to detect TGF-β1 levels in the peripheral blood of patients with lung cancer and 75 healthy volunteers as normal controls.RESULTS AND CONCLUSION: Among the 248 patients, there were 82 BCI-positive cases, and 166 BCI-negative cases.Thirteen patients were confirmed to have catheter-related bloodstream infection. There were 48 Gram-positive bacteria, 24Gram-negative bacilli, and 10 fungal. The levels of TGF-β1 were higher in BCI-positive patients than BCI-negative patients (P < 0.05); the levels of TGF-β1 in the BCI-negative group were higher than those in the normal control group (P < 0.05). For lung cancer patients with nosocomial infection induced BCI, there are various species of pathogenic bacteria, and Gram-positive bacteria are more common. To detect TGF-β1 levels in patients with lung cancer is of significance for early prevention of BCI.

Objective To establish a multiplex PCR targeting the intergenic spacer regions (IGS) for the identification of Cryptococcus neoformans var.grubii,var.neoformans and Cryptococcus gattii.Methods Primers were designed by using the software ClustalW2 and Oligo 6 based on the sequence of IGS1 region,which shows high sequence variability in the genome of Cryptococcus neoformans and Cryptococcus gattii.,for the multiplex PCR.Then,the developed multiplex PCR was performed to identify 51 Cryptococcus neoformans strains representing genotypes VNI-VNIV and VNB as well as 41 Cryptococcus gattii strains representing genotypes VGI-VGIV.The identification results were compared with those from common PCR by using primers GPA1A,CLA4D and SODlgattii specific to Cryptococcus neoformans var.grubii,var.neoformans and Cryptococcus gattii,respectively,as well as with those from the canavanine-glycine-bromothymol blue (CGB) medium-based culture.Results The developed multiplex PCR successfully identified the 92 Cryptococcus neoformans and Cryptococcus gattii.strains,and yielded negative results from the other tested pathogenic yeasts,which revealed a high specificity of the designed primers.False positive results were observed in the identification of two Cryptococcus gattii strains with GPA1A primer-based PCR,one Cryptococcus gattii strain with CLA4D primer-based PCR,one var.grubii strain and one var.neoformans strain with CGB culture,while no false negative results were observed in the detection of these Cryptococcus strains by any of these methods.Conclusions The developed multiplex PCR in this study can rapidly and accurately identify Cryptococcus neoformans var.grubii,var.neoformans,AD hybrid,and Cryptococcus gattii,with superior performance in comparison with common PCR and CGB medium-based culture.

Objective To observe the influences of Bcl-2 expression and pancreatic cancer cell proliferation after downregulating Beclin1 expression and discuss the mechanism.Methods Target interfering plasmid siBeclin1 was constructed and transfected to PANC-1 cell,and cells were divided into non transfected group(Beclin1 group),blank control group(control siNegative group) and transfection group(siBeclin1 group).Western blot and qRT-PCR were used to detect the transfection efficiency and Bcl-2 expression after transfection.Proliferation was detected by MTT.Results Western blot and qRT-PCR results showed that expression of Beclin1 in siBeclin1 group was lower than Beclin1 and siNegative control groups (27.823± 1.432,8.635±1.481,26.904±1.098;F=8.176,P＜0.01),qRT-PCR showed that the Beclin1 mRNA level of PANC-1 cells in siBeclin1 group was lower than Belcin1 and siNegative groups (0.421 ±0.157,0.194±0.104,0.399 ±0.123;F=5.239,P＜0.01),and the silencing rate was about 60％-70％.Bcl-2 protein expression and levels of mRNA in siBeclin1 group were significantly higher than that of Beclin1 group(26.912±1.927,8.004±1.534,t =7.329,P＜0.01;0.582±0.297,0.217±0.186,t =6.835,P＜0.01);MTT results showed that the proliferation of PANC-1 cells were strengthened,(61.54±6.81)％ and (46.78±7.28)％ at 72 h,(76.39±7.26)％ and (54.27±8.17)％ at 96 h,and the difference was significant(t=3.674,P＜0.05;t =10.185,P ＜0.01).Conclusion Proliferation of pancreatic cancer cells are strengthened after Beclin1 expression downregulating,so Beclin1 may inhibit the proliferation of pancreatic cancer cells by Bcl-2 downregulating.Beclin1 can be a target candidate gene of pancreatic cancer therapy.

Objective To evaluate the clinical effect of group cognitive behavioral therapy on blood glucose levels,anxiety and depression in patients with type2 diabetes mellitus.Methods Ninety three patients with type 2 diabetes mellitus were recruited and randomly divided into intervention group (n =48) and control group (n =45).Both groups received diabetes health education,patients in intervention group received additional group cognitive behavioral therapy.The glucose tolerance,glycosylated hemoglobin A1c were measured; the HAMA(Hamilton Anxiety Scale)scores,HAMD(Hamilton Depression Scale)scores and CSQ (Coping Styles Questionnaire) scores in patients were analyzed before and 6 months after treatment.Results After 6-month treatment the fasting blood glucose (6.33 mmol/L vs.5.94 mmol/L),1 h postprandial plasma glucose(12.40 mmol/L vs.11.46 rmool/L),2 h postprandial plasma glucose (10.24 mmol/L vs.9.13 mmol/L),A1 c (6.31％ vs.6.07％) in intervention group were decreased significantly,compared to baseline values (all P ＜ 0.05).The HAMA total score (9.98 vs.8.14),somatic anxiety (3.98 vs.3.48),psychic anxiety(6.00 vs.4.67),HAMD total score(10.74 vs.6.93),anxiety somatic(5.02 vs.3.26),block(2.24 vs.1.38)and sleep disorders(2.40 vs.1.40)in intervention group were all decreased significantly(P ＜ 0.01 or 0.05).There were significant differences in HAMA total score (8.14 vs.9.15),HAMD total score(6.93 vs.9.33),anxiety somatic(3.26 vs.4.38),block(1.38 vs.1.98)and sleep disorders(1.40 vs.2.03)between the intervention group and control group(P ＜ 0.01 or 0.05).And the negative coping style scores in intervention group was also lower than that of the baseline (26.74 vs..29.43).Conclusion The group cognitive behavioral therapy combined with diabetes health education for patients with type 2 diabetes mellitus may improve the glucose metabolism and depression and anxiety status of patients.