Objective To explore the dosage and injection method of concanavalin A(Con A) for inducing Wistar rats into the acute hepatic injury model. Methods (1)According to the dosage of Con A, 42 Wistar rats were randomly divided into groups A, B, C, D, E, N, 7 rats in each group. Group N was given tail intravenous injection of normal saline as normal control group. Groups A, B, C, D, E were given intravenous injection of 4, 8, 16, 30, 40 mg/kg of Con A respectively. At the 8th hour after modeling, the levels of alanine transaminase(ALT), aspartate aminotransferase(AST), albumin(ALB), interleukin(IL)-2 , IL-10, interferon (IFN)-γ, and tumor necrosis factor(TNF)-αwere detected. And HE staining was used to observe the pathological feature of hepatic tissue. (2)According to the injection method of Con A, 21 Wistar rats were randomly divided into normal control group, intraperitoneal injection group and tail intravenous injection group, 7 rats in each group. The dosage of Con A for the rats in intraperitoneal injection group and tail intravenous injection group was 16 mg/kg. At the 8th hour after modeling, the levels of serum ALT, AST, and ALB were determined. Results The number of abnormal deaths in various dose Con A groups at the end of each experiment was 0 in groups A, B, C, and 2 in group D, and 7 in group E. A small amount of spotty necrosis, inflammatory cell infiltration, and hepatic lobule with almost integrity of structure were found in groups A, B, while obvious bridging-like necrosis was seen in groups C, D. Serum ALT, AST, and ALB levels in intraperitoneal injection group had no statistically significant difference as compared with the normal control group. Conclusion Tail intravenous injection of 16 mg/kg of Con A can be used to induce an acute immunological liver injury rat model successfully.

OBJECTIVETo study the effect of the sphingosine kinase 1 (SphK1) inhibitor N,N-dimethylsphingosine (DMS) in combination with chemotherapeutic drugs (DDP, 5-Fu, MMC) on the proliferation of gastric cancer cells (SGC7901) in vitro, and to evaluate whether SphK1 inhibitors could be used as synergetic agents in chemotherapy.

METHODSSGC7901 cells were incubated in vitro with DMS (1 micromol/L) and 5-Fu, DDP, MMC at different concentrations in combination or separately for 24 h. The effects on the growth and survival of SGC7901 cells were determined by MTT assay. The inhibition rates were assessed by response surface analysis and the interactive relationships between the combined drugs were evaluated on the basis of positive/negative values of the cross product coefficients in the response surface equation.

RESULTSThe growth inhibition rate of the gastric cancer cells by treatment with DMS (1 micromol/L) was (10.23 +/- 0.74)%. The growth inhibition rates of the gastric cancer cells treated with 5-Fu (1, 5 and 25 microg/ml) for 24 h were (9.95 +/- 3.24)%, (21.04 +/- 2.19)%, and (45.49 +/- 3.60)%, respectively. The growth inhibition rates of the gastric cancer cells treated with DDP (0.5, 2.5 and 12.5 microg/ml) for 24 h were (9.38 +/- 0.79)%, (19.61 +/- 0.90)%, and (29.83 +/- 0.54)%, respectively. The growth inhibition rates of the gastric cancer cells treated with MMC (0.1, 0.5 and 2.5 microg/ml) for 24 h were (15.35 +/- 0.77)%, (24.72 +/- 0.83)%, and (30.68 +/- 0.28)%, respectively. There were significant differences among the inhibition rates caused by different concentrations of the drugs (P < 0.05). When 1 micromol/L DMS was used in combination with 5-Fu (1, 5, and 25 microg/ml) for 24 h, the growth inhibition rates of the cancer cells were (16.76 +/- 0.41)%, (27.28 +/- 0.29)% and (52.56 +/- 3.60)%, respectively. When 1 micromol/L DMS was used in combination with DDP (0.5, 2.5, and 12.5 microg/ml) for 24 h, the growth inhibition rates of the cancer cells were (15.35 +/- 0.86)%, (25.57 +/- 0.27)%, (36.37 +/- 0.51)%, respectively. When 1 micromol/L DMS was used in combination with MMC (0.1, 0.5, and 2.5 microg/ml) for 24 h, the growth inhibition rates of the cancer cells were (21.02 +/- 0.28)%, (32.10 +/- 0.27)%, (36.36 +/- 0.28)%, respectively. There were also significant differences among the growth inhibition rates caused by different concentrations of the drugs alone and in combination groups (P < 0.05).

CONCLUSIONSDMS can suppress the proliferation of SGC7901 cells in vitro, and there are evident synergetic effects when it is used in combination with chemotherapeutic drugs. The results of this study indicate that SphK1 inhibitors may become novel and promising chemotherapeutic sensitizers.

Antibiotics, Antineoplastic ; pharmacology ; Antimetabolites, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Drug Synergism ; Enzyme Inhibitors ; pharmacology ; Fluorouracil ; pharmacology ; Humans ; Mitomycin ; pharmacology ; Phosphotransferases (Alcohol Group Acceptor) ; antagonists & inhibitors ; Sphingosine ; analogs & derivatives ; pharmacology ; Stomach Neoplasms ; pathology

OBJECTIVEThe purpose of this study was to investigate the correlation between gene mutation and gene copy number and their association with the clinical profiles and pathological features in Chinese patients with non-small cell lung cancer (NSCLC).

METHODSSurgical specimens of cancer tissue were collected from 118 NSCLC patients. Gene mutations in exon 19 and exon 21 were detected by real-time PCR and gene copy number was detected by fluorescence in situ hybridization (FISH). Chi-square (χ(2)) test was performed to analyze the correlation between EGFR mutation and gene copy number, and explore their association with clinicopathological features in the NSCLC patients.

RESULTSThe mutation frequency in EGFR was 41.5% (49/118). EGFR mutations occured in 50.0% (48/96) of patients with adenocarinoma and 5.0% (1/20) of patients with squamous cell carcinoma. EGFR gene high copy number was detected in 70.3% (83/118)of the patients. The FISH-positive rate was 78.1% (75/96) in adenocarcinoma and 35.0% (7/20) in squamous cell carcinoma. EGFR mutation and high copy number mainly occurred in the adenocarcinoma, advanced stage, female gender, and non-smoking patients. There was a significant correlation between EGFR gene mutation and gene high copy number.

CONCLUSIONSEGFR gene mutation and gene high copy number are more common in Chinese NSCLC patients with adenocarcinomas, advanced stage, non-smokers and females. There is a significant correlation between gene mutation and gene high copy number. Combined analysis of EGFR mutation and gene copy number by FISH may provide a superior approach in selecting patients who may benefit from anti-EGFR target therapy.

Adenocarcinoma ; genetics ; pathology ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; Carcinoma, Squamous Cell ; genetics ; pathology ; Exons ; Female ; Gene Dosage ; Genes, erbB-1 ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Mutation ; Mutation Rate ; Neoplasm Staging ; Sex Factors ; Smoking

Capsid ; Cell Line ; Dependovirus ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Hepatitis B virus ; genetics ; Humans ; Liver Diseases ; therapy ; Plasmids ; Recombination, Genetic ; Transfection

Objective To investigate the effects of blue light on the thickness of corneal epithelium and full-thickness of the cornea in mice by optical coherence tomography angiography (OCTA).Methods Totally 40 mice were collected and randomly divided into experimental group and control group,with 20 mice in each group,and the experimental mice were raised in the blue light environment from 8 to 16 hours per day,while the controls were reared in normal environment.Then the thickness of corneal epithelium and full-thickness of the cornea in both groups were measured by OCTA before irradiation and one week,two weeks,one month,two months and three months after irradiation,respectively.Results Compared with pre-irradiation,the thickness of corneal epithelium of all regions did not change significantly in both groups at 1 week,2 weeks,and 1 month after irradiation,and the differences were not statistically significant (all P ＞ 0.05).Compared with before irradiation,the corneal epithelium thickness of the control group at 2 months and 3 months after irradiation did not change significantly,and there was no significant difference (both P ＞ 0.05).Compared with the control group,the corneal epithelium at central,nasal 5 mm,inferior 5 mm,and temporal 5 mm regions in the experimental group were significantly thickened,and the differences were statistically significant (all P ＜0.05).Three months after irradiation,compared with the control group,the thickness of corneal epithelium in the central and inner regions of the cornea and nasal 6 mm and temporal 6 mm regions of the experimental group were significantly thickened,and the differences were statistically significant (all P ＜ 0.05).There was no significant change in the corneal full thickness between the experimental group and the control group before irradiation and 1 week,2 weeks,1 month,2 months,and 3 months after irradiation,and the differences were not statistically significant (all P ＞ 0.05).Furthermore,the difference in the extremum value of corneal epithelial thickness,namely the maximum and the minimum,was significantly different in both groups (P ＜ 0.05),but the difference in the extremum value of the full-thickness of the cornea was not significant in the two groups (P ＞ 0.05).Conclusion The blue light can change the thickness of corneal epithelium in mice,and the change of the central region is obvious,but the full-thickness of the cornea do not significantly change in a short term.

The aim is to observe the expression of human factor VIII gene in mice tranduced in vivo and ex vivo. The vector pLNC-FVIII BD was generated by cloning a B-domain-deleted (760aa-1639aa) FVIII cDNA (FVIIIBD cDNA) into retroviral vector pLNCX. 2 x 10(6) of mouse bone marrow stroma cells transduced by LNC-FVIII BD were infused into 4-week-old BALB/c mice by tail-vein injection. pLNC-FVIII BD was conjugated with PAMAM dendrimer to form complex PAMAM-pLNC-FVIII BD, with which C57BL/6J were injected by tail vein (200 micro l contained 15 micro g/mouse) and sacrificed at days 1, 2, 7, 14, 21 and 28, respectively after injection. Tissue such as liver, spleen, lung and kindney were harvested, with which the transcription were detected by means of RT-PCR. In addition, blood was collected to be measured human FVIII Ag, human FVIIIc and anti-FVIII of human inhibitors. The results showed that the highest level of human FVIII in the recipient BALB/c mice was 8.6 +/- 1.44 ng/ml detected on the first day post-injection; anti-FVIII antibodies were detected from the first week post-injection, and then the level of FVIII Ag decreased and cannot be measured on the fourth week. In the C57BL/6J mice physiological level of human FVIII was expressed in plasma at 48 hours after injection and the average human FVIIIc was 0.62 U/ml and the average human FVIII Ag was 115.5 ng/ml, and gradually reduced later. Anti-FVIII of human inhibitors was not revealed all the time. Syngene image scanning demonstrated that the transcription of the human FVIII BD cDNA occurred mainly in spleen and lung, and secondarily in liver and kidney. No side effects of PAMAM-pLNC-FVIII BD were observed in mice tissue by pathological examination at 4 weeks. In conclusion, retrovirus-transduced bone marrow stroma cells effectively produced human FVIII after ex vivo transduction, but the development of anti-FVIII antibodies in recipient mice influenced the expression level. The human FVIII gene can successfully be transduced in vivo through injecting PAMAM-pLNC-FVIII BD cDNA into mice intravenously. There was physiological level expression of human FVIII in plasma at 48 hours after injection and the average human FVIIIc is 0.62 U/ml and the peak in the six mice was 0.89 U/ml, and gradually reduced later.
Animals ; DNA, Complementary ; analysis ; Factor VIII ; genetics ; Genetic Therapy ; Hemophilia A ; therapy ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Transfection

OBJECTIVE: To explore the role of cell-surface nucleolin in lipopolysaccharide (LPS)-stimulated expression and secretion of TNF-alpha and IL-1beta in human THP-1 monocytes. METHODS: Immuno-fluorescence assay and Western blot were used to identify the expression of nucleolin on the surface of THP-1 monocytes. Inactivation of nucleolin was induced by anti-nucleolin monoclonal antibody blockage, and the expression and secretion of TNF-alpha and IL-1beta were observed by using reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immuno-sorbent assay (ELISA)respectively in LPS-mediated human THP-1 monocyte inflammatory model. RESULTS: Immuno-fluorescence showed that nucleolin was localized on the cell surface of THP-1 monocytes as indicated by dotted red fluorescence. Western blot assay indicated that nucleolin existed in the cell membrane fractions. RT-PCR assay showed that the expressions of TNF-alpha and IL-1beta mRNA significantly increased at 2 h and 3 h after the treatment with 1000 microg/L LPS. After 1 h pretreatment with anti-nucleolin antibody, the levels of TNF-alpha and IL-1beta mRNA decreased compared with an anti-nucleolin antibody untreated group and an irrelevant IgG+LPS group (P<0.05). ELISA assay showed that the pretreatment with anti-nucleolin antibody inhibited significantly the secretion of LPS-induced levels of TNF-alpha and IL-1beta after 4, 12 and 24 h treatment with 1000 microg/L LPS. CONCLUSION Nucleolin expresses on the cell surface of THP-1 monocytes and involves in the LPS-mediated expression and secretion of TNF-alpha and IL-1beta.
Cell Line ; Cell Membrane ; metabolism ; Humans ; Interleukin-1beta ; biosynthesis ; metabolism ; Lipopolysaccharides ; pharmacology ; Monocytes ; cytology ; metabolism ; Phosphoproteins ; metabolism ; physiology ; RNA-Binding Proteins ; metabolism ; physiology ; Tumor Necrosis Factor-alpha ; biosynthesis ; metabolism

Epidermal growth factor receptor (EGFR) gene mutation and copy number are useful predictive markers that guide the selection of non-small cell lung cancer (NSCLC) patients for EGFR-targeting therapy. This study aimed to investigate the correlation between EGFR gene mutation and copy number and clinicopathologic characteristics of Chinese patients with NSCLC. NSCLC specimens collected from 205 patients between November 2009 and January 2011 were selected to detect EGFR gene mutations with real-time polymerase chain reaction (RT-PCR) and to detect EGFR gene copy number with fluorescence in situ hybridization (FISH). EGFR mutations primarily occurred in females, non-smokers, and patients with adenocarinomas (all P < 0.001). Tissues from 128 (62%) patients were FISH-positive for EGFR, including 37 (18%) with gene amplification and 91 (44%) with high polysomy. EGFR gene mutation was correlated with FISH-positive status (R = 0.340, P < 0.001). Multivariate analysis showed that not smoking (OR = 5.910, 95% CI = 2.363-14.779, P < 0.001) and having adenocarcinoma (OR = 0.122, 95% CI = 0.026-0.581, P = 0.008) were favorable factors for EGFR gene mutation. These results show a high frequency of EGFR FISH positivity in NSCLC tissues from Chinese patients and a significant relevance between EGFR gene mutations and FISH-positive status. Among the FISH-positive samples, EGFR gene mutation occurred more frequently in samples with gene amplification compared to those with high polysomy, suggesting that EGFR mutation and gene amplification should be used as clinical decision parameters to predict response to EGFR-targeting therapy.
Adenocarcinoma ; genetics ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; Female ; Gene Amplification ; Gene Dosage ; Humans ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; genetics ; metabolism ; Male ; Middle Aged ; Mutation ; Real-Time Polymerase Chain Reaction ; Receptor, Epidermal Growth Factor ; genetics ; metabolism ; Smoking

Response criteria remain controversial in therapeutic evaluation for locally advanced esophageal carcinoma treated with neoadjuvant chemotherapy. We aimed to identify the predictive value of tumor regression grading (TRG) in tumor response and prognosis. Fifty-two patients who underwent neoadjuvant chemotherapy followed by esophagectomy and radical 2-field lymphadenectomy between June 2007 and June 2011 were included in this study. All tissue specimens were reassessed according to the TRG scale. Potential prognostic factors, including clinicopathologic factors, were evaluated. Survival curves were generated by using the Kaplan-Meier method and compared with the log-rank test. Prognostic factors were determined with multivariate analysis by using the Cox regression model. Our results showed that of 52 cases, 43 (83%) were squamous cell carcinoma and 9 (17%) were adenocarcinoma. TRG was correlated with pathologic T(P = 0.006) and N (P < 0.001) categories. Median overall survival for the entire cohort was 33 months. The 1- and 2-year overall survival rates were 71% and 44%, respectively. Univariate survival analysis results showed that favorable prognostic factors were histological subtype (P = 0.003), pathologic T category (P = 0.026), pathologic N category (P < 0.001), and TRG G0 (P = 0.041). Multivariate analyses identified pathologic N category (P < 0.001) as a significant independent prognostic parameter. Our results indicate that histomorphologic TRG can be considered as an alternative option to predict the therapeutic efficacy and prognostic factor for patients with locally advanced esophageal carcinoma treated by neoadjuvant chemotherapy.
Adenocarcinoma ; drug therapy ; pathology ; surgery ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Squamous Cell ; drug therapy ; pathology ; surgery ; Chemotherapy, Adjuvant ; Esophageal Neoplasms ; drug therapy ; pathology ; surgery ; Esophagectomy ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neoadjuvant Therapy ; Neoplasm Grading ; Organoplatinum Compounds ; administration & dosage ; Predictive Value of Tests ; Proportional Hazards Models ; Retrospective Studies ; Survival Rate ; Taxoids ; administration & dosage

OBJECTIVETo evaluate the relationship between coronary artery stenosis and internal carotid artery, vertebral and basilar artery stenosis using computed tomographic angiography.

METHODSThe imaging and clinical data of 84 patients who underwent coronary, head and cervical computed tomographic angiography in our hospital between September 2008 and June 2010 were retrospectively analyzed. Segment stenosis scoring was performed to quantify the degree of stenosis of coronary arteries. The relationship between the segment stenosis scoring and the degree of stenosis for internal carotid artery, vertebral and basilar artery was analyzed.

RESULTThe coronary segment stenosis scores were significantly correlated with the degree of stenosis for internal carotid artery, vertebral and basilar artery (r=0.450 and 0.475,P<0.05) .

CONCLUSIONThe degree of the stenosis of coronary artery is associated with the stenosis of internal carotid artery, vertebral and basilar artery, which can be confirmed by computed tomographic angiography."

Aged ; Angiography ; methods ; Carotid Artery, Internal ; diagnostic imaging ; Carotid Stenosis ; diagnostic imaging ; Coronary Stenosis ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods ; Vertebrobasilar Insufficiency ; diagnostic imaging