Background: In our previous paper, we demonstrated that Connexin 43 (CX43) was highly expressed in bladder cancer (BC) tissues. But the molecular mechanism about microRNAs (miRNAs) regulation upstream of CX43 in BC has not been well elucidated and remains to be further studied. MicroRNA-139-5p (miR-139-5p) is a tumor suppressor in progression of multifarious cancers including BC. Nevertheless, the underlying mechanisms of CX43/miR-139-5p in tumorigenesis of BC are still not well illustrated. The specific objective of our study was to inquiry the effect of CX43/miR-139-5p on BC progression and its underlying mechanism. Methods: The bioinformatics analysis softwares were applied to predict the miRNAs in the upstream of CX43. First, the expression levels of miR-139-5p in BC tissues (tumor) and paracancer tissues (normal) were investigated using the data from The Cancer Genome Atlas database. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression level of miR-139-5p in three human BC cell lines 5637, T24, ECV-304 and a human bladder epithelial immortalized cell line SV-HUC-1 (normal control). Then si-CX43, si-control, miR-139-5p mimic, and its negative control (NC) were transfected into BC cell line ECV-304. The relationship of miR-139-5p and CX43 was analyzed by dual-luciferase reporter assay. The qRT-PCR and Western blotting were used to test the mRNA and protein expression level of CX43. The proliferation of ECV-304 and T24 cells were examined by cell counting kit-8. The migration and invasion of ECV-304 cells were tested by transwell assay. To determine whether miR-139-5p would affect cell proliferation, migration and invasion by targeting CX43, we executed the rescue assay. The comparison between two groups was analyzed by Student’s t test, and comparisons among multiple samples were performed by oneway analysis of variance and a Bonferroni post hoc test. Results: The expression of miR-139-5p was remarkably down-regulated in BC tissues (tumor vs. normal, 2.286 ± 0.017 vs. 3.211 ± 0.034, t= 11.540, P < 0.0001) and cell lines (P < 0.01 in all BC cell lines). Besides, we also indicated that over-expression of miR-139-5p reduced the proliferation of ECV-304 (P = 0.001) and T24 cells (P = 0.005). Moreover, miR-139-5p over-expression weakened the invasion (P = 0.001) and migration (P = 0.001) of ECV-304 cells. Furthermore, the relative luciferase activity of CX43-wild type construct was distinctly lessened by up-regulation of miR-139-5p (miR-139-5p mimic NC vs. miR-139-5p mimic, 0.916 ± 0.063 vs. 0.356 ± 0.048, t = 7.085, P = 0.002), nevertheless the activity of CX43-mutant type construct was untouched (miR-139-5p mimic NC vs. miR-139-5p mimic, 0.918 ± 0.057 vs. 0.878 ± 0.039, t= 0.577, P = 0.595). Finally, the rescue assay revealed that CX43 deletion enhanced the depressor effect of miR-139-5p on ECV-304 cell proliferation (P < 0.01), invasion (P = 0.028), and migration (P = 0.014). Conclusion MiR-139-5p, as a tumor-suppressor, repressed cell proliferation, invasion, and migration in BC, which might be achieved by regulating CX43.

Background:The increasing prevalence of colorectal cancer (CRC) in China and the paucity of information about relevant expenditure highlight the necessity of better understanding the financial burden and effect of CRC diagnosis and treatment.We performed a survey to quantify the direct medical and non-medical expenditure as well as the resulting financial burden of CRC patients in China.Methods:We conducted a multicenter,cross-sectional survey in 37 tertiary hospitals in 13 provinces across China between 2012 and 2014.Each enrolled patient was interviewed using a structured questionnaire.All expenditure data were inflated to the 2014 Chinese Yuan (CNY;1 CNY =0.163 USD).We quantified the overall expenditure and financial burden and by subgroup (hospital type,age at diagnosis,sex,education,occupation,insurance type,household income,clinical stage,pathologic type,and therapeutic regimen).We then performed generalized linear modeling to determine the factors associated with overall expenditure.Results:A total of 2356 patients with a mean age of 57.4 years were included,57.1％ of whom were men;13.9％ of patients had stage Ⅰ cancer;and the average previous-year household income was 54,525 CNY.The overall average direct expenditure per patient was estimated to be 67,408 CNY,and the expenditures for stage Ⅰ,Ⅱ,Ⅲll,and Ⅳ disease were 56,099 CNY,59,952 CNY,67,292 CNY,and 82,729 CNY,respectively.Non-medical expenditure accounted for 8.3％ of the overall expenditure.The 1-year out-of-pocket expenditure of a newly diagnosed patient was 32,649 CNY,which accounted for 59.9％ of their previous-year household income and caused 75.0％ of families to suffer an unmanageable financial burden.Univariate analysis showed that financial burden and overall expenditure differed in almost all subgroups (P ＜ 0.05),except for sex.Multivariate analysis showed that patients who were treated in specialized hospitals and those who were diagnosed with adenocarcinoma or diagnosed at a later stage were likely to spend more,whereas those with a lower household income and those who underwent surgery spent less (all P ＜ 0.05).Conclusions:For patients in China,direct expenditure for the diagnosis and treatment of CRC seemed catastrophic,and non-medical expenditure was non-ignorable.The financial burden varied among subgroups,especially among patients with different clinical stages of disease,which suggests that,in China,CRC screening might be cost-effective.

OBJECTIVETo discuss the relevant factors of biliary complications after liver transplantation and to investigate the value of comprehensive management for the complications.

METHODSThe data of 366 patients undergoing liver transplantation from October 2000 to March 2012 was analyzed retrospectively, and the risk factors were analyzed by univariate analysis and Stepwise Logistic regression. The cases with biliary leak were administered thorough drainage. The cases with anastomotic biliary stricture were administered sacculus dilatation through percutaneous transhepatic cholangiography (PTC) and endoscopicretrograde cholangiopancreatography (ERCP). If necessary, some cases were placed biliary tract brackets. The patients with nonanastomotic biliary stricture were treated with PTC plus choledochoscope.

RESULTSAll the 366 patients were followed up for 58.5 (10 to 129) months. Biliary complications after liver transplantation were diagnosed in 42 cases among these patients. The incidence for biliary complications was 11.5%. The univariate analysis and multivariate Logistic regression analysis showed that the second warm ischemia period and the blood loss and the damage of blood supply and the diameter of biliary anastmosis were significantly associated with biliary complications after liver transplantations (Wald = 9.474 to 17.208, P < 0.05). Twelve cases with biliary leak were cured through abdominal and nasobiliary drainage. Twenty-two cases with anastomotic biliary stricture were administered sacculus dilatation through ERCP or PTC and were cured, including 6 cases were placed biliary tract brackets. Among 8 cases with nonanastomotic biliary stricture, 6 cases were cured through PTC associating with choledochoscope. One case was treated second liver transplantation and another case got worse.

CONCLUSIONSIschemic injury and the diameter of anastmosis are risk factors for biliary complications after liver transplantations. The interventional management of biliary stricture and bile leakage after liver transplantation is safe and effective.

Adolescent ; Adult ; Aged ; Biliary Fistula ; therapy ; Biliary Tract Diseases ; epidemiology ; therapy ; Cholangiopancreatography, Endoscopic Retrograde ; Female ; Humans ; Liver Transplantation ; Logistic Models ; Male ; Middle Aged ; Postoperative Complications ; epidemiology ; therapy ; Retrospective Studies ; Risk Factors ; Young Adult

LPS stimulation of macrophages production of IFN-beta plays a key role in innate immunity defending the microbial invasion. In this study, the effect of S632A3 promoting LPS-induced IFN-beta production and the underlying mechanism were investigated, mRNA level was measured by real-time PCR, cytokine production was determined by ELISA, GSK-3beta activity was investigated by kinase assay, protein phosphorylation and expression were evaluated by Western blotting. The results revealed that S632A3 significantly augmented IFN-beta production by LPS-stimulated macrophages. S632A3 inhibition of the activation of GSK-3beta, reduced the threonine 239 phosphorylation of transcription factor c-Jun but increased the total level of c-Jun in LPS-stimulated macrophages. Moreover, small interfering RNA-mediated knockdown of c-Jun level abrogated the ability of S632A3 to augment IFN-beta. The study thus demonstrates S632A3 being a new anti-inflammation lead compound and provides a molecular mechanism by which S632A3 promoted LPS-induced IFN-beta production in macrophages through inhibiting the activation of GSK-3beta.
Animals ; Anti-Bacterial Agents ; pharmacology ; Cell Line ; Enzyme Activation ; drug effects ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Interferon-beta ; biosynthesis ; genetics ; Lipopolysaccharides ; pharmacology ; Macrophages ; cytology ; metabolism ; Mice ; Phosphorylation ; Piperidones ; pharmacology ; Proto-Oncogene Proteins c-jun ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo evaluate the value of GeneSearch(TM) BLN assay as an intraoperative diagnostic method of sentinel lymph node metastases in breast cancer patients.

METHODSNinety consecutive patients were involved in this study. SLNs were intraoperatively identified and dissected, and then sectioned vertically to the long axis into multiple blocks. The odd blocks were tested by BLN assay and even ones prepared for frozen sectioning (FS), while all blocks were evaluated by touch imprint cytology (TIC). Post-operatively, residual tissues of the even blocks were assessed by histopathologic examination (4 - 6 µm thick serial sectioning permanent H&E slides were performed every 150 µm and one block made 6 slides).

RESULTSBLN assay could be performed within less than 35 min after learning curve of 10 cases. A correlation was found between cycle time values of mammaglobin or cytokeratin-19 and size of metastases, with Spearman correlation coefficients of 0.67 and 0.71, respectively. The accuracy, sensitivity, specificity, positive predict value (PPV) and negative predict value (NPV) of the assay were 95.6%, 93.3%, 96.7%, 93.3% and 96.7%, While FS had the sensitivity, specificity, PPV, NPV of 76.7%, 100%, 100%, 89.6%, and TIC of 73.3%, 100%, 100%, 88.2%, respectively. The sensitivity of the assay was higher than that of FS (P = 0.07), and was significantly higher than that of FS (P = 0.04). When assessing patients with micro-metastases, the assay had a sensitivity of 85.7%, which was significantly higher than that of FS and TIC (P = 0.03).

CONCLUSIONGeneSearch(TM) BLN Assay can replace FS and TIC for the intraoperative assessment of SLN.

Breast Neoplasms ; diagnosis ; pathology ; Cytodiagnosis ; Frozen Sections ; methods ; Humans ; Keratin-19 ; analysis ; Lymph Nodes ; pathology ; Lymphatic Diseases ; pathology ; Lymphatic Metastasis ; pathology ; Sensitivity and Specificity ; Sentinel Lymph Node Biopsy ; methods

BACKGROUNDSentinel lymph node (SLN) biopsy has become a common procedure for early breast cancer patients. The GeneSearch(TM) Breast Lymph Node (BLN) Assay is a real-time RT-PCR assay for the detecting nodal metastases larger than 0.2 mm. China Breast Cancer Clinical Study Group (CBCSG)-001a is a prospective multi-center clinical trial that was conducted to validate the GeneSearch(TM) BLN Assay in China.

METHODSThe SLNs from 90 consecutive patients were identified and dissected, and then sectioned along the short axis into multiple blocks. Intra-operatively, the odd blocks were tested by BLN assay and the even ones were used for frozen section, while all the blocks were evaluated by touch imprint cytology. Post-operatively, the remaining tissues were assessed by histological evaluation.

RESULTSA total of 189 SLNs was tested by BLN assay. The sensitivity, specificity, positive predictive value, and negative predictive value were 88.9%, 97.4%, 88.9% and 97.4%, respectively, for BLN assay, 75.0%, 100%, 100% and 94.4%, respectively, for frozen section, and 63.9%, 100%, 100% and 92.2%, respectively, for touch imprint cytology. The sensitivity of BLN assay was higher than that of touch imprint cytology (P = 0.01) and frozen section (P = 0.13). When assessing the nodes with micro-metastases, BLN assay had a significant higher sensitivity than frozen section (P = 0.023) and touch imprint cytology (P = 0.005).

CONCLUSIONThe GeneSearch(TM) BLN Assay is an accurate and rapid intra-operative assay for breast SLNs and it is suitable for application in general medical practice.

Adult ; Aged ; Breast Neoplasms ; complications ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; diagnosis ; Middle Aged ; Sentinel Lymph Node Biopsy ; methods

OBJECTIVETo analyze the risk factors for acute renal failure (ARF) early after liver transplantation.

METHODSThe data of 362 patients undergoing liver transplantation from August 2000 to December 2010 were retrospectively analyzed, including 71 patients with ARF (ARF group) and 291 without ARF (non-ARF group). Thirty-six variables, including clinical and experimental variables, were analyzed by t test for continuous variables and χ(2) test for discrete variables. The variables with significance (P < 0.05) were then analyzed with Stepwise logistic regression.

RESULTSTwelve variables, including pretransplant serum creatinine, hemoglobin, thrombinogen activity, total bilirubin, MELD scores, total operation time, intraoperative blood loss, intraoperative blood transfusion, preoperative urine output, preoperative hepatic encephalopathy, intraoperative low blood pressure and postoperative infection, had significant difference between two groups (F = 10.30 - 182.70, P = 0.000 - 0.041). The Stepwise logistic regression analysis for 12 variables demonstrated that the high level of pretransplant serum creatinine, the low pretransplant thrombinogen activity, the high MELD scores, the large volume of intraoperative blood loss, postoperative infection were the independent risk factors of ARF complicated to orthotopic liver transplantation (P < 0.05).

CONCLUSIONSEarly ARF is a key negative factor for the survivors after orthotopic liver transplantation. The reason for ARF complicated to OLT is multiple. The high level of pretransplant serum creatinine, the low pretransplant thrombinogen activity, the high MELD scores, the large volume of intraoperative blood loss, postoperative infection were the independent risk factors of ARF complicated to orthotopic liver transplantation.

Acute Kidney Injury ; etiology ; Adolescent ; Adult ; Aged ; Female ; Humans ; Liver Transplantation ; adverse effects ; Logistic Models ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Retrospective Studies ; Risk Factors ; Young Adult

OBJECTIVETo investigate the value of 11C-PD153035 as an EGFR imaging agent in C6 tumor-bearing rat.

METHODSThe tumor-bearing rats were generated by subcutaneous injection of glioma C6 cells. Positron emission tomography/computer tomography (PET/CT) scans started as soon as intravenous injection of 11C-PD153035 (15-20 MBq/0.3 ml) was completed, images were collected continuously. The region of interest (ROI) was used to study the percentage of radioactivity in major organs and implanted tumors in the rats. The accumulation and blocking study in vitro was completed.

RESULTSThere were significant differences in 11C-PD153035 uptake among major organs. The maximum uptake in the organs ranked in the following order: liver > gastrointestinal tract > kidney > lung > brain > muscle. Radioactivity could be also observed in the tumors. The radioactivity ratio (T/NT, target/non-target) peaked (4.15) at 40 - 50 min post injection. The in vitro blocking study showed that 11C-PD153035 uptaken by C6 cells could be blocked by PD153035.

CONCLUSIONThe results of this study show that 11C-PD153035 can be uptaken by EGFR-expressing tumors. 11C-PD153035 has a potential as a bioprobe to yield useful information for both diagnosis and therapy of tumors. However, the high concentration of 11C-PD153035 in the gastrointestinal tract is unfavorably affecting the tumor detection in these organs.

Animals ; Brain Neoplasms ; diagnostic imaging ; metabolism ; pathology ; Carbon Radioisotopes ; Cell Line, Tumor ; Gastrointestinal Tract ; metabolism ; Glioma ; diagnostic imaging ; metabolism ; pathology ; Liver ; metabolism ; Male ; Neoplasm Transplantation ; Positron-Emission Tomography ; Quinazolines ; pharmacokinetics ; Rats ; Rats, Wistar ; Receptor, Epidermal Growth Factor ; metabolism ; Tissue Distribution ; Tomography, X-Ray Computed

OBJECTIVETo evaluate the effect on increasing bone cement-bone interface micro-gomphosis intensity with bone cement oscillator.

METHODSOne hundred femoral bones of adult pig were randomly divided into 6 groups: oscillating group (A1) and control group (A2) of anti-tensile force, oscillating group (B1) and control group (B2) of anti-pressure (n = 20 in each group), oscillating group (C1) and control group (C2) of imaging (n = 10 in each group). Mechanics and CT test was performed, micro-gomphosis intensity of bone cement-bone interface between oscillating group and control group was compared.

RESULTSMechanics and CT test showed bone cement-bone interface micro-gomphosis intensity in oscillating group was significantly stronger than control group (P < 0.01).

CONCLUSIONBone cement oscillator can significantly increase micro-gomphosis intensity of bone-cement interface, and reduce long-term aseptic loosening of artificial prostheses.

Animals ; Bone Cements ; Cementation ; Equipment Design ; Femur ; Joint Prosthesis ; Male ; Materials Testing ; Mechanics ; Random Allocation ; Swine ; Vibration

OBJECTIVETo explore the awareness on sever acute respiratory syndrome (SARS) and public health emergencies among general publics.

METHODSA cluster sampling method was implemented in Harbin and Jiagedaqi district of Daxinanling of Heilongjiang province. Research subjects were divided into three groups as city, township and rural areas and were given questionnaires to fill in. Data was analyzed with Epi-data and SPSS.

RESULTS2003 available questionnaires were collected. The general publics well understood the knowledge on public health emergencies and the SARS with the whole recognition rate more than 60 percent. During the epidemics, people in city, town and countryside were calm (71.7%). The rates of attitude towards the government were significantly different among the subjects living with the city, town or rural areas. The city group expressed the highest favor to the government and media, 71.8% of them gave the credit on the control of SARS to the effective method taken by the government and 65.0% of them showed that they had enough confidence on the governmental ability of dealing with crises while the countryside group trusted the hospitals and relative specialists the most.

CONCLUSIONIt is essential for the government to interact and communicate with the publics through media, medical and related institutions when confronting with the public health emergencies. Publicity on health knowledge and coping system on emergency should play key roles in the development of an effective public health system while the government should lead the battle.

China ; Data Collection ; Emergencies ; Health Knowledge, Attitudes, Practice ; Humans ; Public Health ; Rural Population ; Severe Acute Respiratory Syndrome ; Urban Population