Objective To investigate whether prophylactic antifungal treatment will reduce the incidence of fungal infection(FI)and the mortality in severe acute pancreatitis(SAP).Methods Forty consecutive patients with SAP admitted from January 2008 to December 2009 were divided into prevention group and control group with 20 cases each by random digits table.The patients in prevention group underwent routine therapy plus intravenous fluconazol as prophylactic antifungal treatment,while routine therapy only in control group.The length of hospital stay,the time when FI occurred,the incidence of FI and mortality were compared with each other.Results Compared with control group,the length of hospital stay wag shorter[(40.3±15.4)d vs.(46.5±18.5)d],the time when FI occurred was slewer[(24.5±14.3)d vs. (14.5±12.7)d],the incidence of FI[15.0%(3/20)vs.35.0%(7/20)]and the mortality [10.0%(2/20)vs. 25.0%(5/20)]were lower in prevention group,there were significant differences between the two groups(P<0.05).Conclusion Prophylactic antifungal treatment can shorten the hospital stay,delay the FI occurrence,reduce the incidence of FI and the mortality in patients with SAP.

Objective To study MRI differential diagnosis of peripheral hemangioma and vascular malformations.Methods MRIfindings of peripheral hemangiomas and vascular malformations proved by operation in 61 cases were retrospectively analyzed in comparison with pathological results.Results In 29 cases of peripheral hemangiomas,20 cases showed equal signal intensity (equal to muscle) and 9 cases showed heterogeneous signal intensity on T1-weighted images; 24 cases showed increased signal intensity (approach fat signal) and 5 cases showed markedly increased signal intensity (increase fat signal) on T2-weighted images,25 cases had septa as low signal network on T2-weighted images,4 cases showed enhancement separated mass enhancement after intravenous contrast injection.In 32 cases peripheral vascular malformations,16 cases showed equal signal intensity and 16 cases showed heterogeneous signal intensity on T1-weighted images.3 cases showed high signal intensity and 29 cases showed markedly increased signal intensity on T2-weighted images,15 cases showed inhomogeneous enhancement after intravenous contrast injection.Conclusion MRI plays an important role in differential diagnosis of peripheral hemangiomas and vascular malformations.

A total of 16 patients with airway stenosis including benign lesion (n=4) and malignant disease (n=12) were treated with argon plasma coagulation (APC) and Z-type covered retrievable metallic stent.L-and I-type stents were placed by guidance of brenchoscope,while Y-type stent was placed by the guidance of both bronchoscope and fluoroscope.Airway stenosis was from (58.8 +9.1)% before APC to (7.5±2.4)% after APC in tracheal,from (67.4±7.4)% to (19.4±4.1)% in left main bronchus,from (69.6±8.9)% to (27.6±5.4)% in right main bronchus.Symptoms of chest distress and breathlessness were improved remarkably,tachypnea indexes were decreased,but Kamofsky performance scope were increased.Twelve stents were successfully installed by the first time,including 9 Y-shape;otherwise,4 stents failed to be installed because the carinal was too wide to insert the Y-shape stent in two patients with lung cancer,finally,L+I type stents were used.Median survival time after successful stenting was 10 months,and mean time was 13 months.APC combined with bifurcated metal stants relieved obstruction and improved quality of life.

The Internet Public Opinion Monitoring and Early Warning System for medical and health industry was designed and implemented due to the frequent occurrence of Internet public opinion, which has all-directional data collecting and analyzing functions, including big data collection, near duplicate detection, spam filtration, key public opinion early warning, region identification and tendency analysis, and can thus provide evidence for relevant departments to take effective measures for the control of Internet public opinion.

Objective To investigate the synergistic therapy effects of B and T lymphocyte attenuator(BTLA) extracellular domain in combination with heat shock protein 70 (HSP70)-TC-1 antigen peptide complex on the mouse model of cervical cancer and the related immunological mechanisms. Methods(1)Detecting the BTLA and herpesvirus entry mediator (HVEM) gene expression in the tumor microenvironment after C57BL/6 mice were inoculated with TC-1 tumor cells by realtime PCR; BTLA,HVEM expression on tumor infiltrating lymphocytes cell surface were detected by flow cytometry (fluorescence intensity). (2) According to different treatments, tumor-bearing mice were divided into 5 groups, which was injected with pcDNA3. 1 (empty vector plasmid as control), psBTLA (vector plasmid which expresses BTLA extracellular domain), HSP70 (HSP70-TC-1 cell peptide complex), HSP70 +pcDNA3.1 or HSP70 + psBTLA, respectively. The weight of tumor was recorded. The expression of immunoregulatory genes in tumor microenvironment were detected. The change of lymphocyte amount and cytotoxicity were detected too; lymphocyte proliferation activity was measured by tritium thymidine incorporation assay; the concentration of interleukin (IL) 2 and interferon-γ(IFN-γ) in supernatants of spleen lymphocyte were measured by enzyme-linked immunosorbent assay (ELISA). Results (1) BTLA gene expression was gradually increased after tumor cells inoculation. The highest expression level was 2. 83 + 0. 35 at 14th day, which had statistical significance difference with the 7th day expression of 1.66±0. 25 (P < 0. 05). While HVEM mRNA expression did not change significantly (P > 0. 05). The 7th and 14th day after TC-1 cells inoculation, the average fluorescence intensity of BTLA expression on the surface of tumor infiltrating lymphocytes was 33.5 and 51.8, respectively, in which there was statistically significant difference (P <0. 05); while the difference of HVEM expression was not statistically significant (57. 2 vs 49. 3 ,P >0. 05). (2)The 28th day after inoculation, tumor inhibition rate of HSP70 + psBTLA group was 88%, which was significantly higher than other treatment groups (P <0. 05). The 28th day after TC-1 cells inoculation, combination therapy not only promoted IFN-γ and IL-2 gene (3. 12 + 0.71,3.20 + 0. 62)expression but also reduced transforming growth factor-β (TGF-β), Foxp3 and IL-10 expression (0. 25±0. 03,0. 19 +0. 03,0. 31 +0. 04;P <0. 05). It also promoted CD8+ T lymphocyte infiltration(52 +6)/high power field, cytotoxicity (65.5±2.4) %, proliferation (15.0 × 103 cpm) and cytokine IL-2 , IFN-γsecretion(824±51), (1096±112) pg/ml, which were all significantly higher than other groups (P <0. 05). Conclusion The effect of immunotherapy on tumor can be augmented by the combination of psBTLA which expresses extracellular domain of BTLA and HSP70-TC-1 tumor antigen peptide complex,which could improve the expression of the related immunoregulatory genes to establish a much better microenvironment in favor of anti-tumor immune response against the mice model of the cervix carcinoma.

Objective To construct eukaryotic expressing vector of mouse soluble CD160 and stably transfect into CHO cells for eukaryotic expression.Methods Recombinant soluble CD160(rsCD160) was constructed by gene recombination.Total RNA was extracted from the spleen of C57BL/6 mice.cDNA was amplified for the soluble form of CD160.Then,the PCR product was cloned tO pcDNA3.1 and pEGFP-N1.The recombinant plasmid was identified by restriction map and sequence analy-sis.The soluble CDl 60 expression in CHO cells transfected with recombinant psCDl 60 was verified by RT-PCR and Westernblot.The binding ability of psCD160 tO its ligand was detected by FACS.Results 520 bp mouse soluble CD160 gene was obtained.Recombinant mouse psCD160 was successfully constructed.After transfection,soluble CD160 expression in the culture supernatant of CHO cells was successfully detected.FACS analysis indicated that soluble CD160 could bind tO its ligand.Conclusion Recombinant mouse psCD160 is successfully constructed,which will benefit our further study on soluble CD160 for immune therapy against tumor in the future experiments.

Objective To explore the effect of blocking BTLA-HVEM (herpesvirus entry mediator-B and T lymphocyte attenuator) pathway on dendritic cell function and the related immunological mechanisms. Methods Murine BTLA extracellular domain eukaryotic expression vector psBTLA was constructed by gene recombination and transfected CHO by Lipofection method. Mouse bone marrow cells were induced to differentiate into DCs by GM-CSF plus IL-4. Expression of BTLA and HVEM on DCs was detected after HSPT0-TC-1 peptide complex stimulation by FACS. Expression of BT-1 and secretion of IL-12 were detected after HSP70-TC-1 peptide complex plus psBTLA transfected CHO culture supernatant stimulation on DCs. Pretreated DCs co-cultured with the same genetic background mouse splenocytes and lymphocytes proliferation and cytokine secretion were detected. Effect of psBTLA gene transfer in vivo on BT-1 expression of DCs and tumor growth on tumor-bearing mice was detected. Results Extracellular domain of murine BTLA was successfully constructed, psBTLA stable transfection CHO cells were obtained and expression of BTLA extracellular domain(sBTLA) was detected the in its culture supernatant. BTLA and HVEM expression of DCs were increased after stimulation by the antigen peptide complex. When DCs were treated with antigen peptide complex plus culture supernatant containing sBTLA, B7-1 expression and IL-12 secretion were increased. Co-cultured with splenocytes, lymphocytes proliferation and cytokine secretion, such as IL-2 and IFN-γ,, were also increased. Gene transfection with psBTLA in vivo promoted B7-1 expression on DCs and inhibited cervical cancer cells growth. Conclusion Blockade of BTLA-HVEM inhibitory pathway with sBTLA can further improve DCs function, activation of lymphocytes and promote antitumor immune response.

Objective To prepare the human papillomavirus (HPV) 16 peptide vaccine and explore the effect in vitro and in vivo. Methods (1) Prediction of the major histocompatibility complex (MHC) class I restricted T cell epitopes by bioinformatics target at transporter associated with antigen processing (TAP) and named by E7Pa, E7Pb, E7Pc separately. (2)In vivo, the C57BL/6 mice were divided into five groups with same amounts randomly after loading with TC-1 cells (HPV 16 positive tumor cells from C57BL/6 mouse), named as E7Pa + CpG,E7Pb + CpG,E7Pc + CpG (as experiment groups, and added 50 μg/ml E7Pa, E7Pb, E7Pc, respectively), CpG(as positive control group and added Con A with 12 mg/L final concentration) and blank control group (without any treatment). The T cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay at different time points;the lactate dehydrogenase (LDH) delivery method was used to test the cytolytie T lymphocyte (CTL) activity of mouse splenic lymphocyte in different ratio of effector cells and target cells (E:T);the related cytokines in tumor tissue and mouse peripheral blood were evaluated by real-time PCR and enzyme-linked immunosorbent assay (ELISA), respectively. The tumor volumes were measured to contrast the therapeutic effect in different groups. Results (1) Three peptide named E7Pa, E7Pb, E7Pc were successfully preparated which had high affinity and specificity. (2) After vaccination of 24, 48, 72,96 hours, MTT results shown that the proliferation rate in E7Pa + CpG group were(131±32)%, (302±15)%, (552±28)%, (731±24)% individually, which were much higher than those in blank control [(72± 15) %, (120 ± 57) %, (176 ±41)%, (288±29)% ;P<0.01], and the other groups i. e. E7Pb + CpG,E7Pc +CpG and CpG groups all proliferated much higher than those in blank control group with statistic signification (P<0. 05), but there was no significant difference between groups(P>0.05);the LDH delivery assay showed that when the ratio of E:T was 100:1, the activity of CTL in the E7Pa + CpG group was most powerful than the other groups with statistic signification (P<0. 01). Meanwhile, the ratio of E:T was concentration-dependent. Compared E7Pb + CpG, E7Pc + CpG or CpG groups with blank control group, there were significantly difference(P<0. 05) ,while there was no significant difference between groups(P >0. 05). The mRNA levels of interferon γ (IFN-γ), interleukin-2 (IL-2) in tumor tissue and peripheral blood in E7Pa + CpG group were significantly higher than those in blank control group (P<0. 01), which was the similar results when compared E7Pb + CpG, E7Pc + CpG or CpG groups with control group (P < 0. 05), and without significant difference between groups(P > 0. 05). The tumor volumes were suppressed obviously in all the experiment groups, especially at the 60th days, the volumes in ETPa + CpG group were much smaller than that in blank control group with statistic signification (P < 0. 01),which was the similar results that E7Pb + CpG, E7Pc + CpG or CpG groups had difference than blank control group with statistic signification (P < 0. 05), and without significant difference between groups(P >0. 05). Conclusion The HPV16 E7 peptide target at TAP combination with CpG as a vaccine could treat effectively the HPV16 E7 positive tumor in experiment.

Objective To investigate the correlation between Young′s elastic modulus (EI) using shear wave elastography (SWE) and liver pathology .Methods Liver biopsy was performed on 231 patients with chronic hepatitis B (CHB) under supersonic guidance ,and SWE with EI of liver was obtained concurrently .The correlation between measured liver stiffness and pathology was analyzed by using the liver pathology as golden standards .One‐way analysis of variance and Spearman rank correlation analysis were performed for the comparison between groups and correlation between two variables , respectively .Receiver operating characteristic (ROC) curve was used to explore the predictive value of shear modulus for the liver inflammation grades and fibrosis stages .Results The EI medians of different liver inflammation grades were 6 .78 kPa (G1) ,7 .30 kPa (G2) ,9 .93 kPa (G3) and 14 .93 kPa (G4) , respectively ,which were statistically different (H=55 .19 ,P<0 .01) .And EI medians of various fibrosis stages were 6 .62 kPa (S0 -S1) ,7 .15 kPa (S2) ,9 .78 kPa (S3) and 14 .62 kPa (S4) ,respectively , which were also significantly different (H=62 .14 ,P<0 .01) .EI was positively correlated with both liver inflammation grades (r=0 .454 6 ,P<0 .01) and liver fibrosis stages (r=0 .505 6 ,P<0 .01) .The areas under the ROC for G≥2 ,G≥3 and G=4 were 0 .68 (95% CI:0 .61 -0 .75) ,0 .77 (95% CI:0 .70 -0 .84) and 0 .85 (95% CI:0 .77-0 .92) ,respectively .The areas under the ROC for S≥2 ,S≥3 and S=4 w ere 0 .73 (95% C I:0 .66 -0 .79 ) ,0 .78 (95% C I:0 .72 -0 .85 ) and 0 .83 (95% C I:0 .75 -0 .91 ) , respectively .Conclusion The EI measured by SWE is correlated with liver pathology of CHB patients , which may be used to dynamically monitor the progress of liver fibrosis .

BACKGROUND AND OBJECTIVETACE, Ar-He target cryosurgery and radioactive seeds implantation are the mainly micro-invasive methods in the treatment of lung cancer. This article summarizes the survival quality after treatment, the clinical efficiency and survival period, and analyzes the advantages and shortcomings of each methods so as to evaluate the clinical effect of non-small cell lung cancer with multiple minimally invasive treatment.

METHODSAll the 139 cases were non-small cell lung cancer patients confirmed by pathology and with follow up from July 2006 to July 2009 retrospectively, and all of them lost operative chance by comprehensive evaluation. Different combination of multiple minimally invasive treatments were selected according to the blood supply, size and location of the lesion. Among the 139 cases, 102 cases of primary and 37 cases of metastasis to mediastinum, lung and chest wall, 71 cases of abundant blood supply used the combination of superselective target artery chemotherapy, Ar-He target cryoablation and radiochemotherapy with seeds implantation; 48 cases of poor blood supply use single Ar-He target cryoablation; 20 cases of poor blood supply use the combination of Ar-He target cryoablation and radiochemotheraoy with seeds implantation. And then the pre- and post-treatment KPS score, imaging data and the result of follow up were analyzed.

RESULTSThe KPS score increased 20.01 meanly after the treatment. Follow up 3 years, 44 cases of CR, 87 cases of PR, 3 cases of NC and 5 cases of PD, and the efficiencywas 94.2%. Ninety-nine cases of 1 year survival (71.2%), 43 cases of 2 years survival (30.2%), 4 cases with over 3 years survival and the median survival was 19 months. Average survival was (16 +/- 1.5) months. There was no severe complications, such as spinal cord injury, vessel and pericardial aspiration.

CONCLUSIONMinimally invasive technique is a highly successful, micro-invasive and effective method with mild complications. To non-small cell lung cancer, we can improve the middle and long term clinical effect by using the different combination of multiple minimally invasive treatments according to the patient's condition.

Carcinoma, Non-Small-Cell Lung ; pathology ; surgery ; Cryosurgery ; methods ; Female ; Humans ; Lung Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome