BACKGROUNDCancer of the esophagus and gastroesophageal junction remains a virulent malignancy with poor prognosis. Rapid progresses were made in chemotherapeutic agents and the development of molecular markers allowed better identification of candidates for targeted therapy. This study aimed to identify the candidate peptides used for anti-angiogenic therapy of esophageal cancer by in vivo screening C7C peptide library for peptides binding specifically to blood vessels of human esophageal cancer.

METHODSThe phage displayed C7C peptide library was injected intravenously into mice bearing human esophageal tumor xenografts under renal capsule. After 5 rounds of screening, 13 clones were picked up individually and sequenced. During each round of screening, titers of phage recovery were calculated from tumor xenograft and control tissues. Homing of these 9 peptides to tumor vessel was detected by calculating phage titers in the tumor xenograft and control tissues (lung and spleen) after each phage was injected into mice model, and compared with the distribution of phage M13 and VIII-related antigen in tumor xenograft by immunohistochemical staining. Comparisons among groups of data were made using one-way analysis of variance (ANOVA), followed by the Bonferroni multiple comparisons test.

RESULTSThe number of phage recovered from tumor tissue of each round increased gradually in tumor group while decreased in control groups (P < 0.01 in tumor and spleen, P < 0.05 in lung). Immunohistochemical staining showed similar staining pattern with M13 antibody or VIII-related antigen antibody, suggesting that phages displaying the selected peptides could home to blood vessel of human esophageal cancer. According to their DNA, 9 corresponding peptide sequences were deduced. And the homing ability to blood vessel of phages displaying the selected peptides was confirmed by comparing with their recovery in tumor and control tissues. Two motifs, YSXNXW and PXNXXN, were also obtained by analyzing the homology of these peptide sequences. The staining distribution of phage with the sequence of PNPNNST was similar to that of the blood vessel marker factor VIII-related antigen staining. After sequencing, each phage with the selected peptide of PNPNNST with 1.0 × 10(11) pfu/ml was injected intravenously into mice. The homing ability to tumor vessel of these 9 kinds of peptides in the xenograft was higher than control tissues (lung and spleen).

CONCLUSIONNine peptides obtained from in vivo screening homed to the blood vessel of human esophageal cancer, and the two motifs of YSXNXW and PXNXXN are the possible biochemical recognition units binding to vascular endothelial cells of esophageal cancer.

Animals ; Antineoplastic Agents ; therapeutic use ; Endothelial Cells ; drug effects ; Esophageal Neoplasms ; blood supply ; drug therapy ; metabolism ; Humans ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Peptide Library ; Peptides ; therapeutic use

OBJECTIVEExperimental evidence shows that microRNAs play an important role in the initiation and progression of human malignancies. The present study aimed to investigate the expressions of 6 microRNAs in prostate cancer (PCa) and their clinical significance.

METHODSWe investigated the expression profiles of 6 microRNAs (let-7g, let-7d, miR-98, miR-96, miR-182 and miR-183) using the method of locked nucleic acid (LNA)-modified oligonucleotide in situ hybridization (ISH) and the technology of tissue microarray (TMA) with the formalin-fixed paraffin-embedded (FFPE) specimens from 52 patients with PCa and 38 with benign prostatic hyperplasia (BPH). Then we analyzed the correlation among the expressions of the 6 microRNAs in PCa and their correlation with the Gleason score and clinical stages of PCa.

RESULTSCompared with BPH, the PCa patients showed decreased expressions of miR-98, let-7d and let-7g, and decreased expressions of miR-96, miR-182 and miR-183, with statistically significant differences between the two groups (P < 0.05). The positive rate of the 6 microRNAs was significantly correlated with the Gleason grades of PCa (P < 0.05), but not with the age and serum PSA concentration of the patients (P > 0.05). The expressions of miR-96 and miR-182 were correlated with the clinical stages of the tumor (P < 0.05). There was a positive correlation among the expressions of miR-96, miR-182 and miR-183 (P = 0.00, r = 0.41), as well as between the expressions of let-7d and let-7g (P = 0.00, r = 0.46) in the PCa tissues. And the expression of miR-98 was positively correlated with those of let-7d and let-7g (P = 0.00, r = 0.46).

CONCLUSIONThe expression profiles of the microRNAs let-7d, let-7g, miR-98, miR-96, miR-182 and miR-183 reflect the biological behavior of PCa to some extent, and might be important biomarkers for the early detection and prognostic assessment of prostate cancer.

Adenocarcinoma ; diagnosis ; metabolism ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; Gene Expression Profiling ; Humans ; Male ; MicroRNAs ; classification ; metabolism ; Middle Aged ; Prognosis ; Prostatic Neoplasms ; diagnosis ; metabolism

ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; antagonists & inhibitors ; genetics ; Adenoviridae ; genetics ; Breast Neoplasms ; therapy ; Cell Line, Tumor ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Gene Silencing ; Humans ; Mitoxantrone ; pharmacokinetics ; Neoplasm Proteins ; antagonists & inhibitors ; genetics ; RNA Interference ; RNA, Small Interfering ; pharmacology

OBJECTIVETo study the postoperative complications and its risk factors in patients underwent breast reconstruction with abdominal flaps.

METHODSThe clinical data of 115 cases underwent breast reconstructions with abdominal flaps from May 2001 to October 2008 was reviewed. The postoperative complications included total flap necrosis, partial flap necrosis, fat necrosis, hernia, bulge, fat liquefaction and infection. The risk factors of complication rates were also evaluated.

RESULTSThe total postoperative complications rate was 17.4% (20/115). No severe complications was found, such as total flap necrosis, hernia and bulge. The most common complications of flap was fat necrosis which occurred in 6 cases (5.2%), partial flap necrosis in 5 cases (4.3%) and infection in 1 case (0.9%). The donor-site complications included fat liquefaction which occurred in 8 cases (7.0%) and infection in 3 cases (2.6%). No significant relation was found between patient's age, body mass index (BMI), timing of surgery and the postoperative complication rate. The postoperative complications occurred more frequently in active smokers, patients with radiotherapy history, or reconstructions with pedicled transverse rectus abdominis myocutaneous (TRAM) flaps. But no significant difference was found in those factors.

CONCLUSIONSFewer complications happens in patients with a reconstruction with deep inferior epigastric perforator (DIEP) flap. Abdominal flap should be performed with more consideration in active smokers or patients with a radiotherapy history. Age and obesity should not be contraindications to breast reconstruction with abdominal flaps.

Abdomen ; surgery ; Adult ; Breast Neoplasms ; surgery ; Female ; Humans ; Mammaplasty ; adverse effects ; methods ; Middle Aged ; Postoperative Complications ; etiology ; Retrospective Studies ; Risk Factors ; Surgical Flaps ; adverse effects

OBJECTIVESThis study is to monitor the survival of E. coli O157:H7 in the aquatic microcosm from Han River and explore the feasibility of fluorescence staining, heterotrophic plate count and ELISA in detection of viable but nonculturable E. coli O157:H7.

METHODSE. coli O157:H7 were added into aquatic microcosm from Han River and cultured with low temperature and oligo-nutrition. Then the survival of E. coli O157:H7 were real-time monitored by acridine orange direct count (AODC), direct viable count (DVC)-CTC staining, DVC-nalidixic acid (NA) staining, heterotrophic plate count (HPC) and ELISA.

RESULTSE. coli O157:H7 can be converted to a viable but nonculturable state in the aquatic microcosm from Han River 58 days after cultured at 4°C with oligo-nutrition. The amount of viable E. coli O157:H7 was measured as 1.2 × 10(5) CFU/ml by DVC-CTC and 9.0 × 10(4) CFU/ml by DVC-NA, whereas the amount of culturable bacterial determined by HPC is 0. The amounts of bacteria determined by ELISA are basically stable within 58 days around 10(6) CFU/ml.

CONCLUSIONE. coli O157:H7 can be converted into a viable but nonculturable state in Han River water at 4°C with oligo-nutrition, and ELISA combined with fluorescence staining and heterotrophic plate count can be used in quantitative detection of the viable but nonculturable E. coli O157:H7.

Colony Count, Microbial ; Culture Media ; Escherichia coli O157 ; isolation & purification ; physiology ; Microbial Viability ; Rivers ; microbiology ; Water Microbiology

This study was aimed to clone the gene coding mouse CXC chemokine receptor 4 (CXCR4), to construct the recombinant lentiviral vector carrying enhanced green fluorescence protein (EGFP) and to explore its expression in eukaryotic cells (293FT cells). The full length CXCR4 gene was cloned by RT-PCR using bone marrow cells from C57BL/6 mouse as template and inserted into PCR-Blunt vector. CXCR4 fragment was generated by digestion with restriction endonuclease and subcloned into a lentiviral vector to generate recombinant lentiviral vector LV-IRES-EGFP-CXCR4, which was co-transfected into 293FT cells together with envelope plasmid and packaging plasmid by lipofectamine 2000. Viruses were gathered and concentrated using ultracentrifuge, and then transfected into 293FT cells. Expression of EGFP was detected by fluorescent microscopy and flow cytometry (FCM). And the expression of CXCR4 protein was detected by Western blot. The results demonstrated that mouse CXCR4 gene was cloned and the lentiviral vector was successfully constructed. The lentiviral particles were correctly packaged, and the virus titers were above 10(8) TU/ml in the supernatant after concentration. Expression of EGFP was detected by fluorescent microscopy in the transfected 293FT cells, and the transfection efficacy > 95% was determined by FCM. Expression of CXCR4 protein detected by FCM and Western blot was significantly higher than that in control group. It is concluded that the CXCR4 gene along with the gene coding EGFP is successfully inserted into a lentiviral vector to construct a recombinant lentiviral vector, which can be expressed in eukaryotic cells.
Animals ; Cell Line ; Gene Expression ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Lentivirus ; genetics ; Mice ; Mice, Inbred C57BL ; Plasmids ; Receptors, CXCR4 ; genetics ; Transfection

OBJECTIVEThe management of secondary normal pressure hydrocephalus (sNPH) is controversial. Many factors may affect the surgery effect. The purpose of this study was to identify the possible factors influencing prognosis and provide theoretical basis for clinical treatment of sNPH.

METHODSA retrospective study was carried out to investigate the results of 31 patients with sNPH who underwent ventriculoperitoneal shunt surgery from January 2007 to December 2011. We processed the potential influencing factors by univariate analysis and the result further by multivariate logistic regression analysis.

RESULTSFactors including age, disease duration and Glasgow coma scale (GCS) score before surgery significantly influenced the prognosis of sNPH (P less than 0.05). Further logistic regression analysis showed that all the three factors are independent influencing factors.

CONCLUSIONAge, disease duration and GCS score before surgery have positive predictive value in estimating favorable response to surgical treatment for sNPH.

Adult ; Female ; Humans ; Hydrocephalus, Normal Pressure ; surgery ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Risk Factors ; Treatment Outcome ; Ventriculoperitoneal Shunt

OBJECTIVETo assess zero drift of intraventricular and subdural intracranial pressure (ICP) monitoring systems.

METHODSA prospective study was conducted in patients who received Codman ICP monitoring in the neurosurgical department from January 2010 to December 2011. According to the location of sensors, the patients were categorized into two groups: intraventricular group and subdural group. Zero drift between the two groups and its association with the duration of ICP monitor were analyzed.

RESULTSTotally, 22 patients undergoing intraventricular ICP monitoring and 27 receiving subdural ICP monitoring were enrolled. There was no significant difference in duration of ICP monitoring, zero drift value and its absolute value between intraventricular and subdural groups (5.38 d+/-2.58 d vs 4.58 d+/-2.24 d, 0.77 mm Hg+/-2.18 mm Hg vs 1.03 mm Hg+/-2.06 mm Hg, 1.68 mm Hg+/-1.55 mm Hg vs 1.70 mm Hg+/-1.53 mm Hg, respectively; all P larger than 0.05). Absolute value of zero drift in both groups significantly rose with the increased duration of ICP monitoring (P less than 0.05) while zero drift value did not. Moreover, daily absolute value in the intraventricular group was significantly smaller than that in the subdural group (0.27 mm Hg+/-0.32 mm Hg vs 0.29 mm Hg+/-0.18 mm Hg, P less than 0.05).

CONCLUSIONThis study demonstrates that absolute value of zero drift significantly correlates with duration of both intraventricular and subdural ICP monitoring. Due to the smaller daily absolute value, ICP values recorded from intraventricular system may be more reliable than those from subdural system.

Aged ; Cerebral Ventricles ; Female ; Humans ; Intracranial Pressure ; Male ; Middle Aged ; Monitoring, Physiologic ; Prospective Studies ; Subdural Space

Objective To analyze time trend of cancer during 1970-2005 in Shandong province so as to develop strategies for control and prevention of cancer at the community level. Methods Data was from 4 retrospective surveys regarding all causes of death during 1970-- 1974, 1985-1989, 1990- 1992 and 2004-2005, in Shandong province. Other than one set of data collected in 1985-1989 by Shandong province itself, the other 3 set of data were from the national surveys, in which the survey-point sampling of choice was based on data of 1970-1974 for assessing its representativeness. The observing indices would include standardized mortality and mortality. A join-point regression model was used to analyze the changing rate of tumor. Results The mortality rate of the entire tumor increased 143.15 percent in 2005 than in 1970. The changing slope of standardized rate of all tumors in the regression model showed that the inter-annual growth rate were 0.54 and 1.24 percent from 1970 to 1984 and from 1985 to 1992. The rate of increase since 1992 had been 0.18 percent. During 2004-2005, the main malignant cancers were lung, stomach, liver, esophageal, coiorectal, leukemia, breast and cervical cancer, in order. Lung cancer rose from the 4th ranking to the first while cervical cancer dropped from the fifth ranking to the 8th place. Esophageal cancer and cervical cancer were decreased annually while gastric cancer was increased in the early days but decreased later on. The rest of the cancers were on the rise year by year. Rates of lung and breast cancers were higher while gastric and esophageal cancers were lower seen in the urban than in rural areas. Conclusion In Shandong province, a marked increase was seen in the mortality rate of tumors in the past 35 years. Evidence showed that the spectrum of death among main malignant tumors had changed which might provide a scientific basis for the development of a community-based prevention and control program on cancer.

OBJECTIVETo investigate the changes of expression of connexin-43 (Cx43) and the tight junction of microvessel endothelial cells (EC) to approach the effects in bleomycin (BLM) induced pulmonary fibrosis (PF).

METHODSForty healthy rats were equally and randomizedly divided into the control group and the experiment group. In both group, vWf in blood serum was measured with ELISA method on 3rd, 7th, 14th, 28th day after BLM treatment. Rats in each group were infused with lanthanum nitrate on 3rd, 7th, 14th, 28th day after BLM treatment. The lung samples were made and the tight junction and the distribution of the granules of lanthanum in the microvessel EC were observed with transmission electron microscopy in the control and experiment groups. The lung microvessel EC of the rats in each group were preserved by tissue culture methods at the same time, and the expression of Cx43 were observed by laser scanning confocal microscopy.

RESULTSThe serum vWf in the peripheral blood of the experiment group was significantly higher than that of the control group, and was the highest on 3rd day after BLM treatment (P < 0.01). The blood vessel EC of the control group were intact. The basement membrane was uninterrupted. Granules of lanthanum nitrate did not penetrate the tight junction of EC. The width of junction gap in the experimental group was increased and the lanthanum granules of high density were found deposited in the linear form in the gap junction. Low expression of Cx43 was observed in experiment group. The expression rate of Cx43 was 25%, 38%, 45% and 71% respectively on 3rd, 7th, 14th, 28th day, significantly less than those in the control group (P < 0.05).

CONCLUSIONIt may be the important pathological basis for the BLM induced abnormality of the interstitial tissues in the lung that the tight junction of EC is continuously in the open state, which causes the effusions of inflammatory cells and the corresponding cytokine secretion, and thus initiates the overproliferation of fibroblasts.

Animals ; Bleomycin ; pharmacology ; Connexin 43 ; biosynthesis ; Endothelial Cells ; drug effects ; metabolism ; Endothelium, Vascular ; cytology ; drug effects ; metabolism ; Female ; Lung ; blood supply ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tight Junctions ; drug effects