ObjectiveTo evaluate the surgical outcomes and prognosis of patients after liver resection for noncolorectal liver metastases.Methods72 patients with liver metastases who underwent liver resection at Tianjin Medical University Cancer Hospital were retrospectively studied.There were 32 males and 42 females,aged between 35～78 years.After liver resection,68 patients had a R0 resection (negative histological margin),and 4 patients had a R1 resection (positive histological margin).The primary tumours were breast,(n =16,22.2 ％),lung (n =14,19.4 ％),gastrointestinal (n=12,16.7％),gynecological (n =8,11.1％),pancreatobiliary (n =8,11.1％),melanoma (n=4,5.6％),sarcoma (n=4,5.6％),and genitourinary (n=2,2.8％).The mean diameter of the main tumour was 4.8 cm (range,1.5- 11.0 cm).The mean number of liver metastases was 1.2 (range,1-5).Liver metastases were synchronous in 6 patients (8.3％) and metachronous in the remaining 66 patients (91.7％).ResultsThe operative mortality was 0％.The mean hospital stay was 14.4 days (range 6-67 days).The median overall survival was 31 months (range,6-127 months).The 1-,3- and 5-year survival rates were 81.9％,37.5％ and 23.6％,respectively.Univariate analysis revealed primary tumour sites (breast vs.others),histological type (adenocarcinoma vs.others),postoperative chemotherapy,number of liver metastases (solitary vs.multiple) and time to liver metastases from diagnosis of primary tumours (≤ 12 months vs.＞ 12 months) were associated with overall survival (all,P＜0.05).In multivariate analysis,factors independently associated with poor survival were nonbreast origin (P =0.012),time to liver metastases from diagnosis of primary tumours ＜12 months (P=0.027) and multiple liver metastases (P=0.008).ConclusionsIn selected patients,liver resection is an effective and safe treatment for noncolorectal liver metastases.The time to liver metastases from diagnosis of primary tumours was independently associated with overall survival.For solitary or liver metastasis of breast origin,surgical resection significantly improved survival.

The effects of linking loop structure between guanine ( G) repeats on G-quadruplex formation were investigated. The results show that the unfavorable effects of long linking loops on G-quadruplex formation can be overcome by introducing double-stranded structures in linking loop regions. This finding provides a new way for sensor design. The activity of G-quadruplex DNAzyme can be tuned by utilizing target-mediated formation of double-stranded structures in loops. As an example, T-T mismatches are introduced in loops to destroy double-stranded structures. The stabilization of Hg2+ to T-T mismatches promotes the reformation of double-stranded structures. Correspondingly, the oligonucleotide folds into G-quadruplex, which binds with hemin to form peroxidase-like G-quadruplex DNAzyme. Hg2+ sensor is designed based on this principle. Using this method, Hg2+ quantitation is achieved in the concentration range of 10-700 nmol/L, with a detection limit of 8. 7 nmol/L. Cysteine will compete with T bases to bind with Hg2+, releasing Hg2+from T-Hg2+-T base pairs. Thus cysteine can also be quantified with this system in the concentration range of 20-700 nmol/L, with a detection limit of 14 nmol/L.

ObjectiveTo investigate the treatment strategies and factors influencing the prognosis of patients with primary gallbladder carcinoma.MethodsThe clinical data of 135 patients with primary gallbladder cancer who were admitted to the Cancer Hospital of Tianjin Medical University from January 2000 to December 2009 were retrospectively analyzed.The survival curve was drawn by the Kaplan-Meier method,and the survival rates were analyzed by using the Log-rank test.Factors which may have influences on the prognosis were analyzed by univariate analysis and COX multivariate analysis.ResultsThe overall 1-,3-,5-year survival rates of the 135 patients were 46.7％,10.4％ and 5.2％,respectively.The 1-,3-,5-year survival rates of 74 patients who received radical resection of gallbladder cancer were 68.9％,18.9％ and 9.5％,respectively.The 1-,3-,5-year survival rates of 50 patients who received palliative treatment were 24.0％,0 and 0,respectively.The 1-,3-,5-year survival rates of 11 patients who received conservative treatment were 0,0 and 0,respectively.There was no significant difference in the survival rates among patients who received different treatment methods (x2 =5.642,P ＜ 0.05 ). Of the 9 patients with gallbladder cancer who received reoperation after laparoscopic choledochotomy,the survival time of 1 patient in stage Ⅰ and 1 of the 3 patients in stage Ⅱ who received radical surgery exceeded 5 years,while the survival time of 5 patients in stage Ⅱ who received palliative treatment was shorter than 5 years.There was a significant difference in the survival time among the 3 groups of patients ( x2 =5.642,P＜0.05).Under the condition of same TNM stages ( Ⅱ,ⅢA,ⅢB,ⅣA,ⅣB),the survival rates of patients who received radical resection of gallbladder cancer were significantly higher than those who received palliative or conservative treatment ( x2 =8.971,21.250,44.153,6.696,21.722,P ＜ 0.05 ).The results of univariate analysis showed that age,CA19-9,TNM stages and treatment methods were risk factors influencing the median survival time ( x2 =8.466,3.977,9.837,5.642,P ＜ 0.05 ).The results of multivariate analysis showed that age,TNM stages and treatment methods were the independent risk factors influencing the median survival time ( Wald=5.779,14.724,11.640,P＜0.05).ConclusionThe prognosis of primary gallbladder cancer is poor.Age,TNM stages and treatment methods are the independent factors influencing the prognosis of patients with gallbladder cancer,and patients who receive radical resection have relatively good prognosis.

Objective:To evaluate the relationship between the mechanism of promoting wound healing by modified autologous blood transfusion and metastasis-associated lung adenocarcinoma transcript 1 ( MALAT1) in diabetic mice. Methods:Twenty SPF ICR mice, weighing 21-25 g, in which the diabetic model was successfully established, were divided into 2 groups ( n=10 each) using a random number table method: modified preservation group (group I) and ordinary preservation group (group O). Peripheral venous blood samples were collected and stored in the corresponding preservation solution for 7 days.The platelet aggregation rate, blood glucose, serum glycosylated hemoglobin (GHB) and phosphodiesterase (DPG) concentrations and WBC were measured.Autologous blood was transfused back immediately after the wound model was established.The percentage of wound healing area was calculated at 7, 10 and 14 days after autologous blood transfusion.The expression of hypoxia-inducible factor-1α, vascular endothelial growth factor, matrix metalloproteinase-9, β-actin, type Ⅰ collagen (Col Ⅰ), Col Ⅲ protein and mRNA and MALAT1 was determined by Western blot, immunohistochemistry and quantitative real-time polymerase chain reaction respectively, at 14 days after transfusion. Results:Compared with group O, the blood glucose, serum concentrations of GHB and DPG, and WBC were significantly decreased, platelet aggregation rate was increased, the percentage of wound healing area was increased, the positive staining rate of Col Ⅰ and Col Ⅲ was increased, and the expression of hypoxia-inducible factor-1α, vascular endothelial growth factor, matrix metalloproteinase-9, ColⅠ, Col Ⅲ and β-actin protein and mRNA and MALAT1 was up-regulated in group I ( P<0.05). Conclusion:The mechanism by which modified autologous blood transfusion promotes wound healing may be related to up-regulating MALAT1 expression in diabetic mice.

Objective To investigate the expression of regulators of G protein signaling(RGS), including RGS2, RGS3 and RGS4 in OLETF rats, as well as the effects of metformin on these expressions. Methods LETO rats were used as control group. Eight-week-old male OLETF rats were assigned to two guoups randomly:model and trial(metfomin dose during 8~(th) to 22~(nd) weeks:300mg kg~(-1)·d~(-1);during 23rd to 28th weeks:400 mg·kg~(-1) ·d~(-1))groups. Expressions of RGS mRNA in aorta and heart werequantified by real-time PCR. Results RGS2, RGS3 and RGS4 mRNA of the thoracic aorta and left ventricle were significantly higher in model group than in control group (P<0.01). Compared with model group, metformin significantly reduced their mRNA in trial group (P<0.01). Conclusions Upregulation of RGS2, RGS3 and RGS4 mRNA expression in the thoracic aorta and left ventricle of OLETF rats is in correlation with cardiovascular lesions; while downregulation of their expression is in correlation with the action of metformin.

Objective:To evaluate the validity and reliability of the Problem Area in Diabetes Scale (PAID) for assessing diabetes-related distress in patients with type 2 diabetes. Methods:Totally 203 outpatients with type 2 diabetes from a tertiary hospital in Beijing were selected. They were assessed with PAID,Hamilton Depression Scale (HAMD-17),Patient Health Questionaire-9(PHQ-9),World Health Organization Five item Well-Being Index (WHO-5 ),Mini-International Neuropsychiatric Interview (MINI)and HbA1 C. Item analysis and exploratory factoranalysis were conducted to test constructive validity. Concurrent validity was evaluated by the correlation coeffi-cients with the other instruments mentioned above. Totally 3 1 subjects were retested 4 weeks later to obtain the test-retest reliability. Results:Exploratory factor analysis produced 4 factors,including emotional,therapeutic,diet and perceived society support problems specific to diabetics. The total variance contribution ratio was 64. 33%. Except i-tem 7 and 20,no item was across two factors. The correlation coefficients of each item with relevant subscale score ranged from 0. 67 to 0. 86. The PAID scores were positively correlated with the scores of HAMD-17,PHQ-9 and HbA1C (r=0. 48,0. 43,0. 21,P<0. 001 or 0. 01),and negatively correlated with WHO-5 scores (r=-0. 46,P<0. 001). The Cronbach's αcoefficients were 0. 94 for the total scale and 0. 81 -0. 88 for the 4 subscales. The test-retest reliability coefficient was 0. 65 for the total scale and 0. 53-0. 73 for the 4 subscales. Conclusion:The validi-ty and reliability of the Problem Area in Diabetes Scale are acceptable in Chinese mainland,and can be available to assess the stress related to diabetics.

Objective:To investigate the effect of different incubation time of aminolevulinic acid (ALA) on photodynamic inhibition of Propionibacterium acnes biofilms. Methods:Propionibacterium acnes biofilms were formed in 24-well plates with pre-placed cell slides and 96-well plates. The formation of the biofilm structure was observed by confocal laser scanning microscopy (CLSM) , and the growth activity of the biofilm was assessed by the tetrazolium salt XTT assay. The in vitro successfully constructed biofilm models were divided into 6 groups: negative control group receiving neither ALA treatment nor LED radiation, ALA group incubated with ALA alone for 30 minutes, LED group receiving LED radiation alone, ALA-PDT1 group, ALA-PDT2 group and ALA-PDT3 group incubated with ALA for 15, 30 and 60 minutes respectively followed by LED radiation. After the treatment, CLSM was performed to observe the biofilm structure, as well as to determine the dead/living bacteria ratio, and XTT assay to assess the growth activity of the biofilm. Differences among groups were analyzed using one-way analysis of variance and least significant difference- t test. Results:CLSM showed that the Propionibacterium acnes biofilm model was successfully constructed in vitro. The dead/living bacteria ratios were 0.90 ± 0.16, 1.75 ± 0.19, and 2.57 ± 0.32 in the ALA-PDT1 group, ALA-PDT2 group and ALA-PDT3 group respectively, which were significantly higher than the dead/living bacteria ratio in the negative control group (0.31 ± 0.01; t= 55.56, 138.62, 74.64, respectively, all P<0.001) ; the biofilm viability value was significantly lower in the ALA-PDT1 group, ALA-PDT2 group and ALA-PDT3 group (0.35 ± 0.02, 0.26 ± 0.02, 0.18 ± 0.01, respectively) than in the negative control group (0.43 ± 0.00; t= 35.66, 2.64, 110.96, respectively, all P < 0.001) . CLSM showed that the structure of the Propionibacterium acnes biofilm was destroyed under the action of ALA-PDT, and the destruction was aggravated with the prolongation of incubation time of ALA. Conclusion:The prolongation of incubation time of ALA can enhance the inhibitory effect of ALA-PDT on Propionibacterium acnes biofilms.

Objective:To investigate the inhibitory effect of deoxyribonuclease Ⅰ (DNaseⅠ) on Cutibacterium acnes biofilms. Methods:Cutibacterium acnes biofilms were constructed, and then were divided into 4 groups (negative control group, 5, 10 and 20 U/ml DNase Ⅰ groups) to be treated with DNase Ⅰ at different concentrations of 0, 5, 10 and 20 U/ml respectively. The biofilm viability was evaluated by tetrazolium salt colorimetric assay, the biofilm content was determined by crystal violet staining-based semi-quantitative analysis, the biofilm structure was observed by confocal laser scanning microscopy, and the live/dead bacteria ratio was calculated. One-way analysis of variance was used to analyze differences between groups. Results:After the treatment with DNase Ⅰ, the biofilm viability was significantly inhibited in the 5, 10 and 20 U/ml DNaseⅠ groups (1.882 ± 0.421, 1.653 ± 0.287, 1.473 ± 0.154, respectively) compared with the negative control group (2.668 ± 0.245), and the inhibitory effect was gradually enhanced with the increase in concentrations of DNase Ⅰ ( F = 9.68, P = 0.005). Crystal violet semi-quantitative analysis showed that the biofilm content was also significantly lower in the 5, 10 and 20 U/ml DNaseⅠ groups (1.039 ± 0.003, 0.489 ± 0.079, 0.147 ± 0.034, respectively) than in the negative control group (1.359 ± 0.071), and the higher the DNase Ⅰ concentration, the lower the biofilm content ( F = 174.40, P < 0.001). Confocal laser scanning microscopy showed that the biofilm structure was destroyed in the 5, 10 and 20 U/ml DNase Ⅰ groups compared with the negative control group, and the higher the DNase Ⅰ concentration, the more severe the destruction of biofilm structure. Additionally, the live/dead bacteria ratio was significantly lower in the 5, 10 and 20 U/ml DNaseⅠ groups (2.303 ± 0.457, 1.534 ± 0.526, 1.263 ± 0.354, respectively) than in the negative control group (4.475 ± 0.146), and the ratio decreased with the increase in concentrations of DNase Ⅰ ( F = 56.75, P < 0.000 1) . Conclusion:DNase Ⅰ had a destructive effect on the structure of Cutibacterium acnes biofilms, and could inhibit their viability.

In the past,we used to think that hepatocellular carcinoma (HCC) only occurred when nonalcoholic fatty liver disease (NAFLD) progressed to liver cirrhosis.However,recent clinical studies have shown that HCC may occur during the stage of nonalcoholic steatohepatitis (NASH).The prevalence of NAFLD and the increasing incidence of NASH-associated HCC require the elucidation of the pathogenesis of NASH-HCC and the assessment of the efficacy of novel therapies as early as possible,while these evaluations need reliable animal models.This article reviews the characteristics of NASH-HCC models with a practical value to uncover the mysteries of NASH-HCC.

Objective:To investigate molecular mechanisms underlying the inflammatory response induced by Cutibacterium acnes ( C. acnes) biofilms in human primary keratinocytes. Methods:A C. acnes biofilm model was established in vitro, and confocal fluorescence microscopy was performed to examine its three-dimensional structure. The cultured human primary keratinocytes were divided into 3 groups: a dimethyl sulfoxide (DMSO) control group (treated with 0.01% DMSO alone), a C. acnes suspension group (co-incubated with C. acnes suspensions), and a C. acnes biofilm group (co-incubated with C. acnes biofilms). Real-time fluorescence-based quantitative PCR (RT-qPCR) was performed to determine the relative mRNA expression of interleukin (IL) -6, IL-8, and tumor necrosis factor (TNF) -α in the groups after 6-hour culture, enzyme-linked immunosorbent assay to detect the free protein levels of IL-6, IL-8, and TNF-α in the groups after 24-hour culture, and Western blot analysis to determine the protein expression of Toll-like receptor 2 (TLR2) in keratinocytes. In addition, some human primary keratinocytes were pretreated with key molecular blockers targeting the TLR2/mitogen-activated protein kinase (MAPK) /nuclear factor (NF) -κB signaling pathway (C29, ST2825, BAY11-7082, SB203580, U0126-EtOH), and then co-incubated with C. acnes biofilms; the DMSO control group and the C. acnes biofilm group receiving no pretreatment were simultaneously set as negative and positive controls, respectively. The mRNA and free protein expression levels of IL-6, IL-8, and TNF-α were then detected in the above groups. One-way analysis of variance was used for comparisons among multiple groups, and the Bonferroni method was used for multiple comparisons. Results:Confocal fluorescence microscopy demonstrated a three-dimensional C. acnes biofilm structure resembling a lawn, and the biofilm grew well. RT-qPCR and ELISA showed significant differences in the mRNA and free protein expression levels of IL-6, IL-8, and TNF-α among the C. acnes biofilm group, C. acnes suspension group and DMSO control group (mRNA: F = 89.70, 312.17, 46.09, respectively, all P < 0.001; free protein: F = 886.12, 634.25, 307.01, respectively, all P < 0.001) ; in detail, the mRNA and free protein expression levels of IL-6, IL-8, and TNF-α were significantly higher in the C. acnes biofilm group than in the C. acnes suspension group and DMSO control group (all P < 0.001) ; the C. acnes suspension group showed significantly increased expression levels of IL-6 mRNA and TNF-α free protein compared with the DMSO control group ( P < 0.001, = 0.003, respectively), while there were no significant differences in the expression of IL-6 free protein, TNF-α mRNA, or IL-8 mRNA and free protein between the 2 groups (all P > 0.05). Western blot analysis showed that the TLR2 protein expression was significantly higher in the C. acnes suspension group and C. acnes biofilm group than in the DMSO control group. After the pretreatment with molecular blockers targeting the MAPK/NF-κB signaling pathway and co-incubation with C. acnes biofilms, the mRNA and free protein expression levels of IL-6, IL-8 and TNF-α were all significantly lower in the C29 group, ST2825 group, BAY11-7082 group, SB203580 group, U0126-EtOH group, as well as in the DMSO control group compared with the C. acnes biofilm group (all P < 0.05) . Conclusion:The C. acnes biofilms exhibited a strong ability to induce inflammatory responses in human keratinocytes, possibly through the activation of the TLR2/MAPK/NF-κB signaling pathway.